TW202035452A - Napi2b-targeted polymer antibody-drug conjugates and methods of use thereof - Google Patents

Abstract

Disclose herein are dosing regimens for targeted NaPi2b antibody-drug conjugates for treating cancer.

Description

Polymer antibody targeting NaPi2b-drug conjugate and method of using it

The present invention generally relates to dosing regimens for administering polymer antibody-drug conjugates targeting NaPi2b to treat cancer. Cross reference of related applications

This application claims the priority and rights of the following U.S. provisional applications under 35 USC § 119(e): No. 62/719,189 filed on August 17, 2018, and No. 62 filed on September 19, 2018 /733,380, No. 62/808,376 filed on February 21, 2019, and No. 62/854,701 filed on May 30, 2019. The complete contents of each of these applications are hereby incorporated for reference. Incorporate the sequence table for reference

The content of the text file named "MRSN-026_001TW_SeqList.txt" created on August 13, 2019 is hereby incorporated for reference.

NaPi2b (SLC34A2, NaPiIIb, Npt2) (multi-transmembrane, sodium-dependent phosphate transporter) (Xu et al. Genomics 62:281-284 (1999)) is normally expressed on the outer brush border membrane of the mammalian small intestine and participates in Inorganic phosphate (Pi) absorption across cells helps to maintain phosphate constant in the body. The protein content of NaPi2b was detected in the liver, on the top surface of breast epithelial cells, salivary glands, and in the lungs, testes, salivary glands, thyroid, small intestine and uterus. NaPi2b mutations have been associated with clinical symptoms of alveolar and testicular microlithiasis. NaPi2b is highly expressed in non-squamous non-small cell lung cancer (NSCLC), non-mucinous ovarian cancer and papillary thyroid cancer. NaPi2b-positive tissue immunoreactivity appeared in 61% of NSCLC and 92% of ovarian cancer samples.

Ovarian cancer is one of the most common gynecological malignancies and the fifth most common cause of cancer death in women. The high mortality rate is partly due to the most common diagnosis of ovarian cancer at an advanced stage, and the mortality rate is approximately 65% of the incidence. Ovarian epithelial tumors account for 58% of all ovarian neoplasms and more than 90% of ovarian malignant tumors. Debulking surgery combined with platinum-based chemotherapy (including taxane) is the current treatment modality; however, most patients with recurrent epithelial ovarian cancer eventually succumb to the disease. There is a need for novel ovarian cancer treatment methods, including targeted therapies, such as immunotherapy with monoclonal antibodies or cancer vaccine-based methods.

NSCLC is any type of epithelial lung cancer other than small cell lung cancer (SCLC). NSCLC accounts for approximately 85% of all lung cancers. Compared with small cell lung cancer, NSCLC is a relatively insensitive category to chemotherapy. When possible, NSCLC is mainly treated by surgical resection for curative intent, although chemotherapy is increasingly used before surgery (leading chemotherapy) and after surgery (adjuvant chemotherapy). Chemotherapy and/or immunotherapy are used in metastatic or non-surgical settings, although most of the disease at this stage is incurable and short survival time remains. There is a need for novel treatments for NSCLC, including targeted therapies, such as immunotherapy with monoclonal antibodies or cancer vaccine-based methods.

Accordingly, there is still a need for treatments targeting the biological activity of NaPi2b.

In various aspects, the present invention provides a method of treating an individual's tumors that express NaPi2b by administering to the individual a fully human or humanized antibody polymer-drug conjugate targeting NaPi2b. The polymer drug conjugate system is administered intravenously at a dose of approximately 10 mg/m ² to 30 mg/m ² on the first day of treatment and every three weeks thereafter. For example, the dose is about 11.5 mg/m ² to about 12.5 mg/m ² , about 19.5 mg/m ² to about 20.5 mg/m ² , about 24.5 mg/m ² to about 25.5 mg/m ² , about 29.5 mg/m ² m ² to about 31.5 mg/m ² , preferably the dose is about 12 mg/m ² or about 30 mg/m ² .

In another aspect, the present invention provides a method of treating an individual’s tumors that exhibit NaPi2b by using a dose of approximately between 10 mg/m ² to 30 mg/m on the first day of treatment and every four weeks thereafter. The target is administered intravenously to the individual by intravenous infusion of doses between m ² , about 10 mg/m ² to 36 mg/m ² or about 10 mg/m ² to 45 mg/m ² NaPi2b is a fully human or humanized antibody polymer-drug conjugate. For example, the dose is about 11.5 mg/m ² to about 12.5 mg/m ² , about 19.5 mg/m ² to about 20.5 mg/m ² , about 24.5 mg/m ² to about 25.5 mg/m ² , about 29.5 mg/m ² m ² to about 31.5 mg/m ² , about 32.5 mg/m ² to about 33.5 mg/m ² , about 35.5 mg/m ² to about 36.5 mg/m ² , about 39.5 mg/m ² to about 41.5 mg/m ^2. About 42.5 mg/m ² to about 43.5 mg/m ² or about 43 mg/m ² to about 45 mg/m ² . Preferably, the dosage is about 30 mg/m ² or about 33 mg/m ² or about 36 mg/m ² or about 40 mg/m ² or about 43 mg/m ² .

Preferably, the dose of intravenous infusion once every four weeks is about 10 mg/m ² , 15 mg/m ² , 20 mg/m ² , 25 mg/m ² , 30 mg/m ² , 33 mg/m ² , 36 mg/m ² , 40 mg/m ² , 43 mg/m ² , 43 mg/m ² , 44 mg/m ² , 45 mg/m ² , or a dose of about 10 mg/m ² , 15 mg/ m ² , 20 mg/m ² , 25 mg/m ² , 30 mg/m ² , 33 mg/m ² , 36 mg/m ² , 40 mg/m ² , or a dose of about 10 mg/m ² , 15 mg/m ² , 20 mg/m ² , 25 mg/m ² , 30 mg/m ² , 33 mg/m ² , 36 mg/m ² , or the dose is about 10 mg/m ² , 15 mg/m ² , 20 mg/m ² , 25 mg/m ² or 30 mg/m ² , 33 mg/m ² , or the dose is about 10 mg/m ² , 15 mg/m ² , 20 mg/m ² , 25 mg/ m ² or 30 mg/m ² , or the dosage is about 10 mg/m ² , 15 mg/m ² , 20 mg/m ² or 25 mg/m ² .

NaPi2b antibody contains an amino acid sequence CDRH1 of GYTFTGYNIH (SEQ ID NO: 5); CDRH2 with amino acid sequence AIYPGNGDTSYKQKFRG (SEQ ID NO: 6); CDRH3 with amino acid sequence GETARATFAY (SEQ ID NO: 7); CDRH3 with amino acid sequence SASQDIGNFLN ( SEQ ID NO: 8) CDRL1; CDRL2 with amino acid sequence YTSSLYS (SEQ ID NO: 9); and CDRL3 with amino acid sequence QQYSKLPLT (SEQ ID NO: 10).

其中： 聚合物包含具有約5 kDa至約10 kDa之分子量範圍的聚(1-羥甲基伸乙基羥甲基-縮甲醛)(PHF)； m為20至75之整數， m₁ 為約5至約35之整數， m₂ 為約3至約10之整數， m_3a 為0至約4之整數， m_3b 為1至約5之整數， m、m₁ 、m₂ 、m_3a 與m_3b 之總和係在約40至約75之範圍內，且 m₅ 為約2至約5之整數。The polymer-drug conjugate contains formula A:

Wherein: The polymer comprises poly(1-hydroxymethylethylene hydroxymethyl-formal) (PHF) with a molecular weight ranging from about 5 kDa to about 10 kDa; m is an integer from 20 to 75, and m ₁ is about An integer from 5 to about 35, m ₂ is an integer from about 3 to about 10, m _3a is an integer from 0 to about 4, m _3b is an integer from 1 to about 5, m, m ₁ , m ₂ , m _3a and m The sum of _3b is in the range of about 40 to about 75, and m ₅ is an integer of about 2 to about 5.

The individual is a human individual. In some aspects, the individual is identified as having NaPi2b performance, as detected by IHC analysis performed on a test cell population obtained from the individual. In other aspects, the individual is identified as having NaPi2b performance, as detected by RNA performance or genetic imprinting in a sample obtained from the individual.

In some aspects, the tumors that express NaPi2b are ovarian cancer, non-small cell lung cancer (NSCLC), papillary thyroid cancer, endometrial cancer, cholangiocarcinoma, papillary renal cell carcinoma, clear cell renal cancer (renal cancer) ), breast cancer, kidney cancer, cervical cancer or salivary duct cancer.

In yet another aspect, the individual has epithelial ovarian cancer, fallopian tube cancer, primary peritoneal cancer, platinum-resistant ovarian cancer, non-squamous NSCLC cancer, (progressive, radioiodine refractory, localized recurrence, or Metastatic disease) papillary thyroid cancer or epithelial endometrial cancer.

In yet another aspect, an individual suffering from epithelial ovarian cancer is sub-classified as high-grade ovarian cancer, low-grade serous ovarian cancer, or clear cell ovarian cancer.

In yet another aspect, an individual with epithelial ovarian cancer has received previous single-dose chemotherapy, such as pegylated liposomal doxorubicin, weekly treatment with paclitaxel, care Topotecan (topotecan), gemcitabine (gemcitabine), PARP inhibitor and the like.

In yet another aspect, an individual with epithelial ovarian cancer has received no more than 3 lines of previous treatment, such as including but not limited to a combination of chemotherapy, such as carboplatin plus paclitaxel, pegylated liposomes and more. Rubicin, weekly treatment with paclitaxel, European paclitaxel (docetaxel), topotecan, gemcitabine, PARP inhibitors and the like. In a further aspect, individuals with NSCLC cancer are subclassified as adenocarcinoma.

In another aspect, the individual has NSCLC and has received previous treatment, such as platinum-based chemotherapy (cisplatin or carboplatin) and PD-1 or PD-L1 monoclonal antibodies. In another aspect, the individual has NSCLC and has received the following previous treatments: carboplatin/paclitaxel, abraxane (nab-paclitaxel), European paclitaxel, pemetrexed (premetrexed), gemcitabine Or a combination of European paclitaxel and ramucirumab.

In another aspect, the individual has NSCLC and has not received additional prior treatment with cytotoxic agents or has not received immunotherapy. In another aspect, individuals with NSCLC have been shown to have intolerance or disease progression to a known cancer-causing mutation and have approved treatments for the mutation (eg, ALK translocation, EGFR mutation).

In a further aspect, a polymer-drug conjugate of Formula A and PD-1 or PD-L1 monoclonal antibody therapy for patients with NSCLC cancer, such as nivolumab, peglizol Mab (pembrolizumab), atezolizumab (atezolizumab), or avelumab (avelumab).

In another aspect, the individual has papillary thyroid cancer that is resistant or intolerant to previous kinase inhibitor therapy or has received previous treatment for low-grade, hormone receptor-positive endometrial adenocarcinoma treatment.

In another aspect, the endometrial cancer is not a stromal tumor or carcinosarcoma.

In another aspect, the individual has endometrial cancer and has received previous treatment with carboplatin/paclitaxel or similar regimens.

In another aspect, the individual has papillary renal cell carcinoma or clear cell renal cell carcinoma, which has a significant papillary growth pattern. In one aspect, the individual has a histological diagnosis of salivary duct cancer that has progressed after standard systemic treatment.

In yet another aspect, the individual is refractory to chemotherapy, including standard first-line chemotherapeutic agents.

In a further aspect, a system of polymer-drug conjugates of Formula A is combined with the following treatments: PARP inhibitors, such as olaparib, niraparib, and lukapa Rucaparib, talazoparib and the like; PD1/PDL-1 inhibitors, such as Nivolumab, Pelizumab, Atezizumab, Aviruzumab and the like Chemotherapy, such as carboplatin, cisplatin, oxaliplatin, doxil, cyclophosphamide, gemcitabine, topotecan, pemetrexe (premetrexe) and the like; VEGF Inhibitors, such as bevacizumab, ramucirumab and the like; tyrosine kinase inhibitors, such as gefitinib, afatinib, erlotinib ( erlotinib), dacomitinib, osimertinib, pazopanib and the like; ALK inhibitors, such as alectinib, crizotinib, Certinib, brigatinib and the like; or BRAF inhibitors such as dabrafenib, trametinib and the like.

In another aspect, the polymer-drug conjugate of formula A is combined with Pelizumab, carboplatin, doxi, bevacizumab, or PARP inhibitor. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those familiar with the technical field of the present invention. In the specification, the singular form also includes the plural form, unless the context clearly dictates otherwise. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, suitable methods and materials are described below. In case of conflict, the present specification (including definitions) will control. In addition, the materials, methods, and examples are only illustrative and not intended to be limiting.

Other features and advantages of the present invention will be apparent from the following detailed description and the scope of patent application.

Detailed description

The present invention provides a method for treating cancers that express NaPi2b by administering a polymer antibody-drug conjugate targeting NaPi2b that specifically binds to the extracellular region of SLC34A2. Specifically, the present invention provides a dosage regimen of XMT-1536 for the treatment of cancers that exhibit NaPi2b. XMT-1536 is composed of about 10 to 15 cysteine moieties conjugated to NaPi2b monoclonal antibody (XMT-1535) via a poly(1-hydroxymethylethylene hydroxymethyl-formal) (PHF) scaffold Auristatin (Auristatin) F-hydroxypropyl amide (AF HPA) conjugate molecules.

Patients with NaPi2b-expressing ovarian cancer , NSCLC , papillary thyroid cancer , endometrial cancer , papillary renal cell carcinoma, or salivary duct cancer were administered XMT-1536 intravenously every three weeks in the dose escalation study . The disease control rate was 67% of patients treated with XMT-1536 at least 12 mg/m ² according to RECIST, version 1.1. XMT-1536 is well tolerated at the maximum dose (40 mg/m ² ) administered. Accordingly, the present invention is characterized by a method for treating tumors exhibiting NaPi2b by administering an infusion dose of about 10 to 45 mg/m ² to the patient (ie, human). Tumors include ovarian cancer , non-small cell lung cancer (NSCLC) , papillary thyroid cancer , endometrial cancer , cholangiocarcinoma , papillary renal cell carcinoma , clear cell renal cancer , breast cancer , kidney cancer , cervical cancer, or salivary ducts cancer. The tumor is a primary tumor or a metastatic tumor.

Patients with NaPi2b-expressing ovarian cancer, NSCLC, papillary thyroid cancer, endometrial cancer, papillary renal cell carcinoma, or salivary gland duct cancer were administered intravenously between about 20 to about every four weeks in the dose escalation study. 45 mg/m ² , that is 20 mg/m ² , 25 mg/m ² , 30 mg/m ² , 33 mg/m ² , 36 mg/m ² , 40 mg/m ² , 43 mg/m ² , XMT-1536 with an infusion dose of 44 mg/m ² or 45 mg/m ² . The individual may or may not have received previous treatment for cancer. For example, the individual has received platinum-based chemotherapy, PD-1 or PD-L1 regimen, or paclitaxel.

In some aspects, the individual is identified as having NaPi2b performance. NaPi2b performance is detected by methods known in the art. For example, with immunohistochemistry (IHC) analysis, fluorescence in situ hybridization (FISH) assay or RNA expression analysis. NaPi2b antibody

The NaPi2b antibody suitable for the method of the present invention specifically binds to the extracellular region of SLC34A2. The present invention further provides a monoclonal antibody targeting NaPi2b, which specifically recognizes NaPi2b, also known as sodium-dependent phosphate transporter 2B. The NaPi2b antibody used in the conjugates disclosed herein can and is useful for modulating, for example, blocking, inhibiting, reducing, antagonizing, neutralizing or otherwise interfering with at least one biological activity of NaPi2b. The antibodies disclosed herein also include antibodies that bind to soluble NaPi2b. The NaPi2b antibody specifically binds to an epitope on the extracellular domain (ECD) of human NaPi2b. These antibodies are collectively referred to herein as "NaPi2b" antibodies.

The NaPi2b antibody-drug conjugate provided herein includes an equilibrium dissociation constant (K _d or K _D ) of ≤1 μM, such as ≤100 nM, preferably ≤10 nM, and more preferably ≤1 nM and NaPi2b epitope Bound antibody. For example, the NaPi2b antibody used in the antibody-drug conjugates disclosed herein exhibits a K _{d in} the range of ≦1 nM to about 1 pM.

The NaPi2b antibody-drug conjugates provided herein may include antibodies capable of modulating, blocking, inhibiting, reducing, antagonizing, neutralizing or otherwise interfering with the functional activity of NaPi2b. The functional activity of NaPi2b includes, for example, participation in transcellular inorganic phosphate (Pi) absorption, thereby helping to maintain phosphate in vivo constant. For example, the NaPi2b anti-system completely or partially inhibits the functional activity of NaPi2b by completely or partially regulating, blocking, inhibiting, reducing, antagonizing, neutralizing or otherwise interfering with transcellular inorganic phosphate absorption. The transcellular inorganic phosphate absorption activity is assessed using any method approved by this technology for detecting transcellular inorganic phosphate absorption activity, including but not limited to the detection of transcellular inorganic phosphate in the presence or absence of the anti-NaPi2b antibody disclosed herein. Phosphate absorption level.

When the functional activity level of NaPi2b in the presence of the NaPi2b antibody is reduced by at least 95%, such as 96%, 97%, 98%, 99% or 100%, compared to the NaPi2b functional activity level in which the NaPi2b antibody described herein does not exist. , Then NaPi2b antibody is considered to completely regulate, block, inhibit, reduce, antagonize, neutralize or otherwise interfere with the functional activity of NaPi2b. When the NaPi2b activity level in the presence of the NaPi2b antibody is reduced by less than 95%, such as 10%, 20%, 25%, 30%, 40%, 50 %, 60%, 75%, 80%, 85% or 90%, NaPi2b antibody is considered to partially regulate, block, inhibit, reduce, antagonize, neutralize or otherwise interfere with the functional activity of NaPi2b.

Exemplary antibodies disclosed herein include XMT-1535 antibody. The antibodies showed specificity for human NaPi2b and showed that the antibodies inhibit NaPi2b activity.

＞XMT-1535重鏈可變區核酸序列

＞XMT-1535輕鏈胺基酸序列(輕鏈可變區(SEQ ID NO：4)(斜體)+ 輕鏈恆定區(SEQ ID NO：12))

＞XMT-1535輕鏈可變區核酸序列

NaPi2b human or humanized monoclonal antibody XMT-1535 includes heavy chain (HC), heavy chain variable region (VH), light chain (LC) and light chain variable region (VL), as shown below And the corresponding nucleic acid sequence. The variable heavy chain region and variable light chain region of each antibody are shaded in the amino acid sequence below. The complementarity determining regions (CDRs) of the heavy and light chains are underlined in the amino acid sequence presented below. The amino acid system containing the complementarity determining region (CDR) of the XMT-1535 antibody is as defined by EA Kabat et al. (see Sequences of Protein of immunological interest, Fifth Edition, US Department of Health and Human Services, Kabat, EA et al. US Government Printing Office (1991)) and disclosed in US Patent 8,603,474. ＞XMT-1535 heavy chain amino acid sequence (heavy chain variable region (SEQ ID NO: 3) (italic) + IgG1 heavy chain constant region (SEQ ID NO: 11))

＞XMT-1535 heavy chain variable region nucleic acid sequence

＞XMT-1535 light chain amino acid sequence (light chain variable region (SEQ ID NO: 4) (italic) + light chain constant region (SEQ ID NO: 12))

＞XMT-1535 light chain variable region nucleic acid sequence

The present invention also includes antibodies that bind to the same epitope as the antibodies described herein or cross-compete with the same epitope as the antibodies described herein. For example, the antibody disclosed herein specifically binds to NaPi2b, wherein the antibody binds to an epitope of one or more amino acid residues on human NaPi2b (such as GenBank Accession No. 095436.3) .

The antibody disclosed herein specifically binds to an epitope on full-length human NaPi2b containing the following amino acid sequences:

The antibodies disclosed herein specifically bind to epitopes on the extracellular domain (ECD) of human NaPi2b.

Those familiar with the art should recognize that it is possible not to use excessive experiments to determine whether a monoclonal antibody has the same specificity as the monoclonal antibody disclosed herein (for example, XMT-1535, 10H1.11.4B), which is based on Determine whether the former prevents the latter from binding to natural binding partners or other molecules known to associate with NaPi2b. If the monoclonal antibody system to be tested competes with the monoclonal antibody disclosed herein, as shown by the reduced binding of the monoclonal antibody disclosed herein, the two monoclonal antibody systems bind to the same or closely related epitopes.

An alternative method to determine whether a monoclonal antibody has the specificity of the monoclonal antibody disclosed herein is to pre-incubate the monoclonal antibody disclosed herein with soluble NaPi2b (which has normal reactivity) and then add the monoclonal antibody to be tested To determine whether the ability of the monoclonal antibody to be tested to bind to NaPi2b is inhibited. If the monoclonal antibody to be tested is inhibited, it is very likely to have the same or functionally equivalent epitope specificity as the monoclonal antibody disclosed herein.

The screening of monoclonal antibodies disclosed herein can also be, for example, by measuring the activity mediated by NaPi2b and determining whether the test monoclonal antibody can regulate, block, inhibit, reduce, antagonize, neutralize or otherwise interfere with NaPi2b activity. get on.

The antibody disclosed herein contains a heavy chain variable region and a light chain variable region, the heavy chain variable region having a sequence selected from the group consisting of SEQ ID NO: 3 at least 90%, 91%, 92%, The amino acid sequence of 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher identity, and the light chain variable region has an amino acid sequence selected from SEQ ID NO: 4 The sequence of the group is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more identical to the amino acid sequence.

In some embodiments, the antibody disclosed herein contains at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% of the amino acid sequence of SEQ ID NO:1. %, 99% or more identical heavy chain amino acid sequence and at least 90%, 91%, 92%, 93%, 94%, 95%, 96% with the amino acid sequence of SEQ ID NO: 2 , 97%, 98%, 99% or higher identity of the light chain amino acid sequence.

The antibody disclosed herein contains a heavy chain variable region and a light chain variable region, the heavy chain variable region having a sequence selected from the group consisting of SEQ ID NO: 3 at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher identity of amino acid sequences, and the light The chain variable region has a sequence selected from the group consisting of SEQ ID NO: 4 at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94% , 95%, 96%, 97%, 98%, 99% or higher identity amino acid sequence.

In some embodiments, the antibody disclosed herein contains an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93% of the amino acid sequence of SEQ ID NO:1. %, 94%, 95%, 96%, 97%, 98%, 99% or higher identity heavy chain amino acid sequence and at least 85%, 86% with the amino acid sequence of SEQ ID NO: 2 , 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher identity light chain amine group Acid sequence.

In some embodiments, the antibody disclosed herein contains the amino acid sequence of the heavy chain variable region of SEQ ID NO: 3 and the amino acid sequence of the light chain variable region of SEQ ID NO: 4.

In some embodiments, the antibody disclosed herein contains the heavy chain amino acid sequence of SEQ ID NO:1 and the light chain amino acid sequence of SEQ ID NO:2.

In some embodiments, the antibody disclosed herein contains the CDRH1 amino acid sequence of SEQ ID NO: 5, the CDRH2 amino acid sequence of SEQ ID NO: 6, the CDRH3 amino acid sequence of SEQ ID NO: 7, and the The CDRL1 amino acid sequence of ID NO: 8, the CDRL2 amino acid sequence of SEQ ID NO: 9 and the CDRL3 amino acid sequence of SEQ ID NO: 10.

In some embodiments, the antibodies disclosed herein contain: at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, and amino acid sequence GYTFTGYNIH (SEQ ID NO: 5) 97%, 98%, 99% or higher identity of the amino acid sequence CDRH1; containing at least 90%, 91%, 92%, 93%, 94 with the amino acid sequence AIYPGNGDTSYKQKFRG (SEQ ID NO: 6) %, 95%, 96%, 97%, 98%, 99% or higher identity of the amino acid sequence CDRH2; containing at least 90%, 91% with the amino acid sequence GETARATFAY (SEQ ID NO: 7) , 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher identity of the amino acid sequence CDRH3; containing the amino acid sequence SASQDIGNFLN (SEQ ID NO: 8) of at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher identity of amino acid sequence CDRL1; contains at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher with the amino acid sequence YTSSLYS (SEQ ID NO: 9) CDRL2 of the amino acid sequence of identity; and containing at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, and the amino acid sequence QQYSKLPLT (SEQ ID NO: 10) CDRL3 with 98%, 99% or higher identity of amino acid sequence.

In some embodiments, the antibody disclosed herein contains: at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, and amino acid sequence GYTFTGYNIH (SEQ ID NO: 5) 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher identity of the amino acid sequence CDRH1; contains the amino acid sequence AIYPGNGDTSYKQKFRG (SEQ ID NO: 6) at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% , 99% or more identical amino acid sequence CDRH2; containing at least 85%, 86%, 87%, 88%, 89%, 90%, and amino acid sequence GETARATFAY (SEQ ID NO: 7) 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher identity of the amino acid sequence CDRH3; containing the amino acid sequence SASQDIGNFLN (SEQ ID NO: 8) At least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or CDRL1 of the amino acid sequence of higher identity; contains at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92 with the amino acid sequence YTSSLYS (SEQ ID NO: 9) %, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher identity of the amino acid sequence CDRL2; and containing the amino acid sequence QQYSKLPLT (SEQ ID NO: 10 ) At least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher The CDRL3 of the amino acid sequence of identity.

In certain embodiments, the antibodies disclosed herein include one or more conservative amino acid substitutions in the variable domain sequence, such as 1, 2, 3, 4, 5, 6, 7, 8, 9 , 10, 11, 12, 13, 14, 15, or more conservative substitutions in the variable domain sequence. In some embodiments, the conservative amino acid substitutions are in the CDR region, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 Or multiple conservative substitutions are accumulated in all CDRs, and in some special embodiments, up to 1, 2, 3, or 4 conservative amino acid substitutions can be present in each CDR sequence, such as SEQ ID NO: 5 to 10.

Those familiar with the technical field should recognize that it is possible not to use excessive experiments to determine whether a monoclonal antibody has the same specificity as the monoclonal antibody XMT-1535, by determining whether the former prevents the latter from binding to a natural binding partner or has Know other molecules associated with NaPi2b. If the monoclonal antibody system to be tested competes with the monoclonal antibody disclosed herein, as shown by the reduced binding of the monoclonal antibody disclosed herein, the two monoclonal antibody systems bind to the same or closely related epitopes.

An alternative method to determine whether a monoclonal antibody has the specificity of the monoclonal antibody disclosed herein is to pre-incubate the monoclonal antibody disclosed herein with soluble NaPi2b (which has normal reactivity) and then add the monoclonal antibody to be tested To determine whether the ability of the monoclonal antibody to be tested to bind to NaPi2b is inhibited. If the monoclonal antibody to be tested is inhibited, it is very likely to have the same or functionally equivalent epitope specificity as the monoclonal antibody disclosed herein.

The screening of monoclonal antibodies disclosed herein can also be, for example, by measuring the activity mediated by NaPi2b and determining whether the test monoclonal antibody can regulate, block, inhibit, reduce, antagonize, neutralize or otherwise interfere with NaPi2b activity. get on.

NaPi2b antibodies suitable for use in the methods disclosed herein can be produced and purified by well-known techniques, such as WO 2009/097128, WO 2017/160754 and US 16/136,706, each of which is incorporated herein in its entirety for reference . Polymer antibody drug conjugate targeting NaPi2b

The present invention relates to treatments involving immunoconjugates comprising antibodies conjugated via polymer scaffolds with cytotoxic agents, such as toxins (for example, enzyme-active toxins or fragments of bacteria, fungi, plant or animal origin) .

The conjugates described herein include NaPi2b antibodies attached to one or more AF-HPA-carrying polymer scaffolds, which independently comprise poly(1-hydroxymethyl) with a molecular weight ranging from about 5 kDa to about 10 kDa Ethylene hydroxymethyl-formal) (PHF). The polymer scaffold carrying AF-HPA is conjugated with the antibody targeting NaPi2b via the cysteine residues of NaPi2b.

其中： 聚合物包含具有約5 kDa至約10 kDa之分子量範圍的聚(1-羥甲基伸乙基羥甲基-縮甲醛)(PHF)； m為20至75之整數， m₁ 為約5至約35之整數， m₂ 為約3至約10之整數， m_3a 為0至約4之整數， m_3b 為1至約5之整數， m、m₁ 、m₂ 、m_3a 與m_3b 之總和係在約40至約75之範圍內， m₅ 為約2至約5之整數，且 NaPi2b為本文所述之全人類或人源化NaPi2b抗體XMT1535。Specifically, the polymer antibody-drug conjugate targeting NaPi2b is XMT-1536 and has the formula (A):

Wherein: The polymer comprises poly(1-hydroxymethylethylene hydroxymethyl-formal) (PHF) with a molecular weight ranging from about 5 kDa to about 10 kDa; m is an integer from 20 to 75, and m ₁ is about An integer from 5 to about 35, m ₂ is an integer from about 3 to about 10, m _3a is an integer from 0 to about 4, m _3b is an integer from 1 to about 5, m, m ₁ , m ₂ , m _3a and m The sum of _3b is in the range of about 40 to about 75, m ₅ is an integer from about 2 to about 5, and NaPi2b is the fully human or humanized NaPi2b antibody XMT1535 described herein.

In some embodiments, m is an integer from about 30 to about 75.

In some embodiments, m is an integer from about 30 to about 40.

In some embodiments, m ₁ is an integer from about 10 to about 20.

In some embodiments, m ₁ is an integer from about 10 to about 12.

In some embodiments, m ₂ is an integer from about 3 to about 5.

In some embodiments, m _3a is an integer from 0 to about 1.

In some embodiments, m _3b is an integer from 2 to about 4.

In some embodiments, m ₅ is an integer from about 2 to about 4.

In some embodiments, m ₅ is an integer from about 3 to about 4.

In some embodiments, the polymer antibody-drug conjugate targeting NaPi2b contains 10 to 15 AF-HPA molecules.

In some embodiments, the PHF has a molecular weight range of about 6 kDa to about 8 kDa.

In some embodiments, the PHF has a molecular weight range of about 6 kDa to about 7 kDa.

其中： m為30至約35之整數， m₁ 為8至約10之整數， m₂ 為2至約5之整數， m_3a 為0至約1之整數， m_3b 為1至約2之整數， m_3a 與m_3b 之總和係在1至約4之範圍內，且 在PHF與抗體之間的比為約3至約5。In a specific embodiment, the NaPi2b-targeted polymer antibody-drug conjugate formula (A) has formula (B), wherein the polymer is PHF with a molecular weight ranging from about 5 kDa to about 10 kDa:

Wherein: m is an integer from 30 to about 35, m ₁ is an integer from 8 to about 10, m ₂ is an integer from 2 to about 5, m _3a is an integer from 0 to about 1, m _3b is an integer from 1 to about 2 , The sum of m _3a and m _3b is in the range of 1 to about 4, and the ratio between PHF and antibody is about 3 to about 5.

The polymer antibody-drug conjugate (ie XMT-1536) that is suitable for use in the method disclosed herein can be produced and purified by well-known techniques, such as WO 2009/097128, WO 2017/160754, PCT/ US18/38988 and US 16/136,706, each of which is incorporated herein in its entirety for reference. Dosage and administration

The cancer treatments provided herein containing polymer antibody-drug conjugates targeting NaPi2b are administered in an amount sufficient to exert a useful therapeutic effect. The active agent is usually administered in an amount that does not cause undesirable side effects in the patient to be treated or minimizes or reduces the side effects observed. Carcinomas that exhibit NaPi2b include, for example, ovarian cancer, non-small cell lung cancer (NSCLC), papillary thyroid cancer, endometrial cancer, cholangiocarcinoma, papillary renal cell carcinoma, clear cell renal cancer, breast cancer, kidney cancer, and cervical cancer Cancer and salivary duct cancer.

Determining the precise amount of active agent (including polymer antibody-drug conjugate targeting NaPi2b) to be administered to an individual is within the skill of those skilled in the art. For example, these agents and uses for the treatment of cancer and solid tumors are well known in the art. Therefore, the dosage of these agents is selected on the basis of the standard dosing schedule of the agent given the route of administration.

It should be understood that the precise dose and duration of treatment are a function of the tissue or tumor to be treated, and can be determined empirically using known test plans or inferred from in vivo or in-vitro test data and/or can be determined from the specific dose. Known dosing schedule is determined. It should be noted that the concentration and dosage value may also vary with the age of the individual being treated, the individual's weight, the route of administration, and/or the degree or severity of the disease and other factors considered within the scope of the physician's skill level. The dosage regimen is usually chosen to limit toxicity. It should be noted that the attending physician will know how and when to terminate, interrupt or adjust the treatment to a lower dose due to toxicity or bone marrow, liver or kidney or other tissue dysfunction. Conversely, if the clinical response is insufficient (excluding toxic side effects), the attending physician will also know how and when to adjust to a higher level. It should be further understood that the specific dosage regimen for any particular individual should be adjusted over time according to the individual needs and the professional judgment of the person who administers or supervises the administration of the formulation, and the concentration range presented in this article is merely illustrative and not intended Limit its scope.

For example, a polymer antibody-drug conjugate system targeting NaPi2b is administered in a therapeutically effective amount to reduce tumor volume.

The amount of polymer antibody-drug conjugate targeting NaPi2b is administered to treat diseases or conditions, such as cancer or solid tumors, and the amount can be determined by standard clinical techniques. In addition, in vitro assays and animal models can be used to help identify the optimal dose range. The precise dose that can be determined empirically may depend on the route of administration, the type of disease to be treated, and the severity of the disease.

The conjugate system provided herein is administered intravenously. The conjugate for intravenous administration can be administered by bolus injection or infusion, infusion, or through a combination thereof. The infusion time may be about 1 minute to 3 hours, such as about 1 minute to about 2 hours, or about 1 minute to about 60 minutes, or at least 10 minutes, 40 minutes, or 60 minutes.

The dosage is between about 10 mg/m ² and 30 mg/m ² . For example, the dosage is between about 11.5 mg/m ² to about 12.5 mg/m ² . Alternatively, the dosage is between about 19.5 mg/m ² and about 20.5 mg/m ² , between about 24.5 mg/m ² and about 25.5 mg/m ² , between about 29.5 mg/m 2 ² to about 31.5 mg/m ² . In some embodiments, the dosage is about 12 mg/m ² , about 20 mg/m ² , about 25 mg/m ² or about 30 mg/m ² . In other implementation aspects, the dosage is 10 mg/m ² , 11 mg/m ² , 12 mg/m ² , 13 mg/m ² , 14 mg/m ² , 15 mg/m ² , 16 mg/m ² , 17 mg/m ² , 18 mg/m ² , 19 mg/m ² , 20 mg/m ² , 21 mg/m ² , 22 mg/m ² , 23 mg/m ² , 24 mg/m ² , 25 mg/m ² , 26 mg/m ² , 27 mg/m ² , 28 mg/m ² , 29 mg/m ² or 30 mg/m ² . In these embodiments, the dose is administered intravenously every four weeks, that is, a 28-day cycle.

Alternatively, the dosage is between about 10 mg/m ² to 33 mg/m ² . For example, the dosage is between about 11.5 mg/m ² to about 12.5 mg/m ² . Alternatively, the dosage is between about 19.5 mg/m ² and about 20.5 mg/m ² , between about 24.5 mg/m ² and about 25.5 mg/m ² , between about 29.5 mg/m 2 ² to about 31.5 mg/m ² or between about 32.5 mg/m ² to about 33.5 mg/m ² . In some embodiments, the dosage is about 12 mg/m ² , about 20 mg/m ² , about 25 mg/m ² , about 30 mg/m ² or about 33 mg/m ² . In other implementation aspects, the dosage is 10 mg/m ² , 11 mg/m ² , 12 mg/m ² , 13 mg/m ² , 14 mg/m ² , 15 mg/m ² , 16 mg/m ² , 17 mg/m ² , 18 mg/m ² , 19 mg/m ² , 20 mg/m ² , 21 mg/m ² , 22 mg/m ² , 23 mg/m ² , 24 mg/m ² , 25 mg/m ² , 26 mg/m ² , 27 mg/m ² , 28 mg/m ² , 29 mg/m ² , 30 mg/m ² , 31 mg/m ² , 32 mg/m ² or 33 mg /m ² . In these embodiments, the dose is administered intravenously every four weeks, that is, a 28-day cycle.

Alternatively, the dosage is between about 10 mg/m ² to 36 mg/m ² . For example, the dosage is between about 11.5 mg/m ² to about 12.5 mg/m ² . Alternatively, the dosage is between about 19.5 mg/m ² and about 20.5 mg/m ² , between about 24.5 mg/m ² and about 25.5 mg/m ² , between about 29.5 mg/m 2 Between ² and about 31.5 mg/m ² , between about 32.5 mg/m ² and about 33.5 mg/m ² or between about 35.5 mg/m ² and about 36.5 mg/m ² . In some embodiments, the dosage is about 12 mg/m ² , about 20 mg/m ² , about 25 mg/m ² , about 30 mg/m ² , about 33 mg/m ² or about 36 mg/m ² . In other implementation aspects, the dosage is 10 mg/m ² , 11 mg/m ² , 12 mg/m ² , 13 mg/m ² , 14 mg/m ² , 15 mg/m ² , 16 mg/m ² , 17 mg/m ² , 18 mg/m ² , 19 mg/m ² , 20 mg/m ² , 21 mg/m ² , 22 mg/m ² , 23 mg/m ² , 24 mg/m ² , 25 mg/m ² , 26 mg/m ² , 27 mg/m ² , 28 mg/m ² , 29 mg/m ² , 30 mg/m ² , 31 mg/m ² , 32 mg/m ² , 33 mg /m ² , 34 mg/m ² , 35 mg/m ² or 36 mg/m ² . In these embodiments, the dose is administered intravenously every four weeks, that is, a 28-day cycle.

Alternatively, the dosage is between about 10 mg/m ² to 40 mg/m ² . For example, the dosage is between about 11.5 mg/m ² to about 12.5 mg/m ² . Alternatively, the dosage is between about 19.5 mg/m ² and about 20.5 mg/m ² , between about 24.5 mg/m ² and about 25.5 mg/m ² , between about 29.5 mg/m 2 between about ² to 31.5 mg / m ^2, between about 32.5 mg / m ² to about 33.5 mg / m ^2, between about ² between 35.5 mg / m ² to about 36.5 mg / m, between about 39.5 mg/m ² to about 41.5 mg/m ² . In some embodiments, the dosage is about 12 mg/m ² , about 20 mg/m ² , about 25 mg/m ² , about 30 mg/m ² , about 33 mg/m ² , about 36 mg/m ² Or between about 40 mg/m ² . In other implementation aspects, the dosage is 10 mg/m ² , 11 mg/m ² , 12 mg/m ² , 13 mg/m ² , 14 mg/m ² , 15 mg/m ² , 16 mg/m ² , 17 mg/m ² , 18 mg/m ² , 19 mg/m ² , 20 mg/m ² , 21 mg/m ² , 22 mg/m ² , 23 mg/m ² , 24 mg/m ² , 25 mg/m ² , 26 mg/m ² , 27 mg/m ² , 28 mg/m ² , 29 mg/m ² , 30 mg/m ² , 31 mg/m ² , 32 mg/m ² , 33 mg /m ² , 34 mg/m ² , 35 mg/m ² , 36 mg/m ² , 37 mg/m ² , 38 mg/m ² , 39 mg/m ² , 40 mg/m ² . In these embodiments, the dose is administered intravenously every four weeks, that is, a 28-day cycle.

Alternatively, the dosage is between about 10 mg/m ² to 45 mg/m ² . For example, the dosage is between about 11.5 mg/m ² to about 12.5 mg/m ² . Alternatively, the dosage is between about 19.5 mg/m ² and about 20.5 mg/m ² , between about 24.5 mg/m ² and about 25.5 mg/m ² , between about 29.5 mg/m 2 between about ² to 31.5 mg / m ^2, between about 32.5 mg / m ² to about 33.5 mg / m ^2, between about ² between 35.5 mg / m ² to about 36.5 mg / m, between about Between 39.5 mg/m ² and about 41.5 mg/m ² , between about 42.5 mg/m ² and about 43.5 mg/m ² , between about 43 mg/m ² and about 45 mg/m ² . In some embodiments, the dosage is about 12 mg/m ² , about 20 mg/m ² , about 25 mg/m ² , about 30 mg/m ² , about 33 mg/m ² , about 36 mg/m ² , About 40 mg/m ² , about 43 mg/m ² or about 45 mg/m ² . In other implementation aspects, the dosage is 10 mg/m ² , 11 mg/m ² , 12 mg/m ² , 13 mg/m ² , 14 mg/m ² , 15 mg/m ² , 16 mg/m ² , 17 mg/m ² , 18 mg/m ² , 19 mg/m ² , 20 mg/m ² , 21 mg/m ² , 22 mg/m ² , 23 mg/m ² , 24 mg/m ² , 25 mg/m ² , 26 mg/m ² , 27 mg/m ² , 28 mg/m ² , 29 mg/m ² , 30 mg/m ² , 31 mg/m ² , 32 mg/m ² , 33 mg /m ² , 34 mg/m ² , 35 mg/m ² , 36 mg/m ² , 37 mg/m ² , 38 mg/m ² , 39 mg/m ² , 40 mg/m ² , 41 mg/m ^2. 42 mg/m ² , 43 mg/m ² , 44 mg/m ² or 45 mg/m ² . In these embodiments, the dose is administered intravenously every four weeks, that is, a 28-day cycle.

The frequency, timing, and dosage of the administration can be administered periodically through the administration cycle to maintain the continuous and/or long-term effect of the active agent for a desired length of time. The provided polymer antibody-drug conjugate composition targeting NaPi2b can be administered hourly, daily, weekly, monthly, yearly or once. The length of the administration cycle can be determined empirically and depends on the disease to be treated, the severity of the disease, the particular patient, and other considerations within the skill level of the treating physician. The length of treatment with the combination therapy provided herein can be one week, two weeks, one month, several months, one year, several years or more.

For example, the frequency of administration of the polymer antibody-drug conjugate targeting NaPi2b is once a day, every other day, twice a week, once a week, once every 2 weeks, once every 3 weeks, or once every 4 weeks. The dosage can be divided into multiple administration cycles during the course of treatment. For example, the polymer antibody-drug conjugate targeting NaPi2b can be administered at a frequency of about 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, one year or more. Give. The frequency of dosing can be the same throughout the cycle or can be different. For example, an exemplary dosage frequency is at least the first week of the administration cycle and twice a week. After the first week, the frequency can continue twice a week, can be increased to more than twice a week, or can be reduced to no more than once a week. The determination of the specific dosage frequency and administration period based on the specific dose to be administered, the disease or condition to be treated, the severity of the disease or condition, the age of the individual, and other similar factors is within the scope of the skilled person.

If the symptoms of the disease persist without stopping the treatment, the treatment can be continued for an additional length of time. Evidence of disease and/or treatment-related toxicity or side effects can be monitored during the course of treatment.

The administration cycle of the polymer antibody-drug conjugate targeting NaPi2b can be adjusted to increase the period of stopping treatment, so as to provide a rest period from exposure to the agent. The length of time for stopping treatment may be a predetermined time or may be determined empirically depending on how the patient responds or depending on the side effects observed. For example, treatment can be stopped for one week, two weeks, three weeks, one month, or several months. The discontinuation period is usually established as the patient's dosing regimen cycle.

An exemplary dosing schedule is a 21-day or 28-day treatment cycle or administration cycle. The dosage regimen is preferably a 28-day treatment cycle or administration cycle. The polymer antibody-drug conjugate system targeting NaPi2b disclosed herein was administered on the 1st day, followed by no administration for 20 days, or on the 1st day, and no administration for the subsequent 27 days. It is within the skill of the technician to determine the precise dosing cycle and schedule.

As mentioned above, the casting cycle can be any length of time desired. Therefore, the 21-day dosing cycle or the 28-day dosing cycle can be repeated for any length of time. For example, a 21-day administration cycle or a 28-day administration cycle can be repeated for 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, and 10 months , 11 months, 12 months, 1.5 years, 2 years, 2.5 years, 3 years or more. Depending on the specific personal considerations of the patient to be treated and the disease, it is within the scope of the skilled person to adopt a dosage cycle and dosage regimen that meets the needs of the patient. Measuring NaPi2b performance

In various aspects, the present invention provides methods for identifying cancer patients who are compliant with targeted NaPi2b therapy or monitoring treatment regimens by measuring the NaPi2b performance status in tumor samples obtained from patients.

In some embodiments, the NaPi2b diagnostic test can be used to identify individuals treated with polymer drug conjugates that target NaPi2b.

The sample is derived from an individual suffering from cancer. The cancer cell sample is excised from tissue removed or obtained from the individual. In some embodiments, the sample is a fresh, frozen or archived biopsy sample.

In some embodiments, the test cell population is derived from fresh unfrozen tissue of the biopsy sample. In other embodiments, the test cell population is derived from the primary or metastatic site. In some embodiments, the test cell population is derived from fresh or frozen tissue or ascites or pleural fluid from a biopsy or surgical sample. In some embodiments, the test cell population is derived from fixed tissues of biopsy or surgical samples (for example, formalin-fixed or formalin-fixed paraffin embedding (FFPE)) or cell masses obtained from fluid samples. Tissue samples can be frozen or fresh.

The necessary NaPi2b performance level can be a level identified by any method known in the art and more specifically by the methods described herein. For example, the expression level of NaPi2b can be measured by performing known immunological assays, such as enzyme immunoassays, radioimmunoassays, competitive immunoassays, double antibody sandwich assays that use antibodies that specifically recognize NaPi2b to express protein expression. Method, fluorescence immunoassay, ELISA, western blotting technique, agglutination assay, cytofluorescence (e.g. flow cytometry), fluorescence in situ hybridization (FISH), colorimetric or immunohistochemical staining assay (IHC). It is particularly desirable to use cell-based assays to determine NaPi2b performance status, such as flow cytometry (FC), immunohistochemistry (IHC), RNA performance analysis, or immunofluorescence (IF), because these assay formats are clinically suitable of.

Flow cytometry (FC) can be used to determine the cell surface expression of NaPi2b in tumor samples before, during and after drug treatment. For example, if so required, flow cytometry can be used to analyze the NaPi2b expression of tumor cells and identify markers such as cancer cell types. Flow cytometry can be performed according to standard methods. See, for example, Chow et al., Cytometry (Communications in Clinical Cytometry) 46: 72-78 (2001). Simply by way of example and illustration, the following planning cytometry analysis: Cells order of 2% paraformaldehyde fixed for 10 minutes at 37 ^o C, followed by 90% of methanol on ice permeabilized for 30 minutes. The cells can then be stained with NaPi2b specific antibody, washed and labeled with a fluorescently labeled secondary antibody. The cells can then be analyzed on a flow cytometer (such as Beckman Coulter FC500) according to the specific plan of the instrument used. This analysis will identify the level of NaPi2b expressed in the tumor.

Immunohistochemistry (IHC) staining can also be used to determine the appearance of NaPi2b in tumor samples before, during and after drug treatment. IHC can be performed according to well-known techniques. See, for example, antibodies; A LABORATORY MANUAL, Chapter 10, Harlow & Lane Eds., Cold Spring Harbor Laboratory (1988). Briefly and by way of example, the paraffin-embedded tissue for histochemical staining (such as tumor tissue from biopsy) is prepared as follows: the tissue section is deparaffinized with xylene and then deparaffinized with ethanol; in water , Then hydrate in PBS; de-mask the antigen by heating the slice in sodium citrate buffer; incubate the slice in hydrogen peroxide; block in the blocking solution; make the slice in the primary peptide antibody and secondary Cultivate in antibodies; and finally use the ABC avidin/biotin method to detect according to the manufacturer's instructions.

Immunofluorescence (IF) assay can also be used to determine the performance in NaPi2b tumor samples before, during and after drug treatment. IF can be performed according to well-known techniques. See, for example, J. M. Polak and S. Van Noorden (1997) INTRODUCTION TO IMMUNOCYTOCHEMISTRY, 2nd Ed.; ROYAL MICROSCOPY SOCIETY MICROSCOPY HANDBOOK 37, BioScientific/Springer-Verlag. Briefly and by way of example, the patient sample can be fixed in paraformaldehyde, then fixed with methanol to block with a blocking solution (such as horse serum), incubated with anti-peptide primary antibodies, and then treated with fluorescent dye The labeled secondary antibody (such as Alexa 488) is incubated and analyzed with an epifluorescent microscope.

The antibodies used in the above assays can be advantageously combined with fluorescent dyes (e.g. Alexa488, PE) or other markers for multi-parameter analysis together with other signal transduction (phospho-AKT, phospho-Erk 1/2) ( Such as quantum dots) and/or cell markers (cytokeratin) antibody conjugates.

In a preferred embodiment, the expression of NaPi2b in tumor-derived samples is determined by immunohistochemistry. In another embodiment, the expression of NaPi2b in a sample from a tumor is determined using the immunohistochemistry (IHC) method described in US 16/136,706, the entire content of which is incorporated herein for reference.

Alternatively, the assay method may include preparing RNA from the sample for PCR (polymerase chain reaction) or other analysis methods as needed. The PCR method is, as necessary, RT-PCR (Reverse Transcription-PCR) or quantitative PCR, such as real-time RT-PCR, RNA seq, and the like. Alternatively, the assay can be performed by using arrays, such as microarrays known in related fields such as nanostring technologies.

The patient is identified as responsive to treatment, where the treatment or detection of cancer is monitored by detecting and/or measuring the performance level of NaPi2b in the sample.

The detection/measurement of NaPi2b performance level is determined by calculating the NaPi2b score. The NaPi2b score is quantitative or semi-quantitative. For example, the testing department uses pathology to score points to obtain pathology scores. It is expected that any scoring method known in the art can be used in the method of the present invention. In particular, any histological scoring method known in the art.

Methods for evaluating measurement results obtained by the histochemical staining assay method include, for example, the H-score method. The H-score is determined by the following calculation formula (Am J Clin Pathol. 1988; 90(3): 233-9). H-score=((% of <1+)X 0)+((% of 1+)X 1)+((% of 2+)X 2)+((% of 3+)X 3), where staining intensity 0 is undyed; staining intensity 1 is weak staining; staining intensity 2 is moderate staining; staining intensity 3 is strong staining.

In the evaluation by the H-score method, only the cancer cell part is used. Formalin-fixed paraffin-embedded cell lines or xenografts (cell lines whose protein expression levels are known in advance) can be used as negative or positive controls for staining intensity. When there is no control sample, multiple samples are evaluated at the same time to confirm the overall distribution of the dyeing intensity of the sample, and then the dyeing intensity can be set.

In addition to the H-score method, other scoring methods known in the art can also be used, such as the Allred method (Harvey et al. Journal of Clinical Oncology 17, No. 5 (May 1999) 1474-1474). Each method requires a cut-off point. Allred score = score of percentage of positive cells + staining intensity score.

The present invention also provides kits and/or methods for identifying or otherwise subdividing (e.g. stratifying) patient groups suitable for therapeutic administration of the antibody-drug conjugates targeting NaPi2b disclosed herein, It is by identifying the individual's NaPi2b score before treatment with the NaPi2b-targeting antibody-drug conjugate disclosed herein. In some embodiments, the test cell population is derived from fresh unfrozen tissue of the biopsy sample. In some embodiments, the test cell population is derived from the primary or metastatic site. In some embodiments, the test cell population is derived from frozen tissue or ascites or pleural fluid of a biopsy or surgical sample. In some embodiments, the test cell population is derived from a fixed tissue (for example, formalin fixation) of a biopsy or surgical sample. The IHC test measures the amount of NaPi2b receptor protein on the cell surface of cancer tissue samples. definition

Unless otherwise defined, the scientific and technical terms used in the present invention shall have the meanings commonly understood by those who are generally familiar with the technical field. Furthermore, unless otherwise required by the context, singular terms shall include pluralities, and plural terms shall include the singular. Regarding the nomenclature used in cell and tissue culture, molecular biology and protein and oligonucleotide or polynucleotide chemistry and hybridization as described herein, and these techniques are generally known and frequently used in the art. Standard techniques are used for recombinant DNA, oligonucleotide synthesis and tissue culture and transformation (e.g. electroporation, lipofection). The enzymatic reaction and purification techniques are carried out according to the manufacturer's instructions or as commonly used implementation methods of this technique or as described herein. The aforementioned techniques and procedures are generally performed according to conventional methods well known in the art and described in various general and more specific documents cited and discussed throughout this specification. See, for example, Sambrook et al., Molecular Cloning: A Laboratory Manual (2d ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (1989)). The nomenclature used in analytical chemistry, synthetic organic chemistry, medical and medicinal chemistry and the laboratory procedures and techniques described herein are those familiar and frequent users in this technology. Standard techniques are used for chemical synthesis, chemical analysis, pharmaceutical preparation, compounding, and patient delivery and treatment.

It should be understood that the following terms used in accordance with the present invention have the following meanings, unless otherwise indicated:

The term “NaPi2b” as used herein (also known as sodium-dependent phosphate transporter 2B, SLC34A2, NaPiIIb, Npt2, Na(+)-dependent phosphate cotransporter 2B, sodium/phosphate cotransporter 2B, Na( +)/Pi cotransporter 2B, NaPi3b, 34 members of the solute carrier family 2) as used herein refers to human NaPi2b (e.g., gene database access code 095436.3) and includes natural expression and expression by cells (including tumor cells) Any variant, homotype and species homologue of NaPi2b on cells transfected with NaPi2b gene. These terms are synonymous and can be used interchangeably.

The term "NaPi2b antibody" or "anti-NaPi2b antibody" as used herein is an antibody that specifically binds to the extracellular region of SLC34A2.

When two or more antibodies are used herein, the term "competition" or "cross-competition" indicates that two or more antibodies compete for binding to NaPi2b, for example, compete for binding to NaPi2b in any assay recognized by the technology. If the antibody competes with one or more other antibodies by 25% or more, the antibody "blocks" or "cross-blocks" the binding of one or more other antibodies to NaPi2b, with 25% to 74% representing "partial blocking". And 75% to 100% means "complete blocking", as measured by any test method approved by this technology. For some antibody pairs, competition or blockade in any of the assays recognized by the technology is only observed when one antibody is applied to the dish and the other antibody is used to compete, and vice versa. Unless otherwise defined or denied in the context, the terms "competition", "cross-competition", "blocking" or "cross-blocking" are also intended to encompass these antibody pairs when used herein.

The term "antibody" as used herein refers to immunoglobulin molecules and immunologically active portions of immunoglobulin (Ig) molecules, that is, molecules that contain an antigen binding site that specifically binds to (immunely reacts with) an antigen. "Specific binding" or "immune response" or "directed against" means that the antibody reacts with one or more antigenic determinants of the desired antigen and does not react with other polypeptides or has a lower affinity (K _d > 10 ^-6 ) Combine. Antibodies include but are not limited to multiple strains, single strains and chimeric antibodies.

It is known that the basic antibody structural unit comprises a tetramer. Each tetrameric system consists of two identical pairs of polypeptide chains, each pair having a "light" chain (about 25 kDa) and a "heavy" chain (about 50 to 70 kDa). The amino terminal portion of each chain includes a variable region of about 100 to 110 or more amino acids that is mainly responsible for antigen recognition. The carboxy terminal part of each chain defines the constant region that is mainly responsible for effector functions. Antibody molecules obtained from humans generally differ from each other in terms of the nature of the heavy chain present in the molecule with respect to any of the classes IgG, IgM, IgA, IgE, and IgD. Certain classes also have subclasses, such as IgG ₁ , IgG ₂ and others. In addition, the light chain in humans can be a kappa (kappa) chain or a lambda (lambda) chain.

As used herein, the term "monoclonal antibody" (mAb) or "monoclonal antibody composition" refers to a group of antibody molecules containing only one molecular type of antibody molecule, which is composed of unique light chain gene products and unique Of heavy chain gene products. In particular, the complementarity determining region (CDR) of a monoclonal antibody is the same in all molecules of the population. A mAb contains an antigen binding site that can immunoreact with a specific epitope of an antigen characterized by its unique binding affinity.

Antibody molecules obtained from humans generally differ from each other in terms of any of the classes IgG, IgM, IgA, IgE, and IgD depending on the nature of the heavy chain present in the molecule. Certain classes also have subclasses, such as IgG ₁ , IgG ₂ and others. In addition, the light chain in humans can be a kappa chain or a lambda chain.

The term "antigen binding site" or "binding portion" refers to the part of an immunoglobulin molecule that participates in antigen binding. The antigen binding site is formed by amino acid residues in the N-terminal variable ("V") region of the heavy ("H") chain and light ("L") chain. The three highly divergent stretches (called ``highly variable regions'') in the V regions of the heavy and light chains are inserted in the more reserved ones known as the ``framework region'' or ``FR''. The flanks extend between. Therefore, the term "FR" refers to the naturally-found amino acid sequence between and adjacent to the hypervariable regions in immunoglobulins. In an antibody molecule, the three hypervariable regions of the light chain and the three hypervariable regions of the heavy chain are arranged opposite to each other in a three-dimensional space to form an antigen-binding surface. The antigen binding surface is complementary to the three-dimensional surface of the bound antigen, and the three hypervariable regions of each of the heavy chain and the light chain are called "complementarity determining regions" or "CDRs". The allocation of amino acids in each domain is in accordance with Kabat Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, Md. (1987 and 1991)) or J. Mol. Biol. 196:901-917 ( 1987), Chothia et al. Nature 342:878-883 (1989) definition.

The term "epitope" as used herein includes any protein determinant capable of specifically binding to immunoglobulins or fragments thereof or T cell receptors. The term "epitope" includes any protein determinant capable of specifically binding to immunoglobulins or T cell receptors. Epitope determinants are usually composed of chemically active surface groupings of molecules (such as amino acids or sugar side chains) and usually have specific three-dimensional structural characteristics, as well as specific charge characteristics. When the dissociation constant is ≤1 μM; for example, ≤100 nM, preferably ≤10 nM, and more preferably ≤1 nM, the antibody is said to specifically bind to the antigen.

The term "polypeptide" is used herein as a general term, which refers to natural proteins, fragments or analogs of multiple peptide sequences. Therefore, natural protein fragments and analogs belong to the genus Polypeptide. The term "naturally occurring" as used herein for objects refers to the fact that objects can be found in nature. For example, polypeptides or polynucleotide sequences that exist in organisms (including viruses) that can be isolated from natural sources and have not been intentionally modified by personnel or other means in the laboratory are naturally produced.

Use the following terms to describe the sequence relationship between two or more polynucleotide or amino acid sequences: "reference sequence", "comparison window", "sequence identity", "sequence identity percentage" and "substantial identity" ". A "reference sequence" is a defined sequence used as a basis for sequence comparison. The reference sequence can be a subset of a larger sequence, for example, as a segment of the full-length cDNA or gene sequence given in the sequence listing, or it can include the complete cDNA or gene sequence. The reference sequence is usually at least 18 nucleotides or 6 amino acids in length, often at least 24 nucleotides or 8 amino acids in length, and often at least 48 nucleotides or 16 amino acids in length . Because two polynucleotide or amino acid sequences can respectively (1) contain similar sequences between the two molecules (that is, a part of the complete polynucleotide or amino acid sequence), and (2) can additionally contain A sequence that diverges between two polynucleotide or amino acid sequences, so sequence comparison between two (or more) molecules is usually performed by comparing the sequences of two molecules through a "comparison window" , To identify and compare the sequence similarity of local regions. The "comparison window" as used herein refers to at least 18 consecutive nucleotide positions or a conceptual segment of 6 amino acids, wherein the polynucleotide sequence or the amino acid sequence can be at least 18 Comparison of the reference sequence of 1 consecutive nucleotide or 6 amino acid sequences, and the polynucleotide sequence part in the comparison window may contain 20% or less when compared with the reference sequence (not including additions or deletions) The additions, deletions, substitutions and the like (ie gaps) make the two sequences optimally arranged. The optimal sequence arrangement of the arrangement comparison window can be obtained by Smith and Waterman's Adv. Appl. Math. 2: 482 (1981) local homology algorithm (local homology algorithm), Needleman and Wunsch's J. Mol. Biol. 48:443 (1970) homology alignment algorithm, Pearson and Lipman's Proc. Natl. Acad. Sci. (USA) 85: 2444 (1988) similarity method search, these algorithms Computer execution (GAP, BESTFIT, FASTA and TFASTA (Genetics Computer Group, 575 Science Dr., Madison, Wis.), Geneworks or MacVector package software in the Wisconsin Genetics package software version 7.0) or inspection, and choose to The best arrangement produced by various methods (that is, the highest homology percentage obtained through the comparison window).

The term "sequence identity" means that two polynucleotide or amino acid sequences are identical through a comparison window (ie, based on nucleotide-to-nucleotide or residue-to-residue). The term "percent sequence identity" is calculated by comparing two optimized permutations through a comparison window, and determining that the same nucleic acid base (such as A, T, C, G, U or I) or residue appears in In the two sequences, the number of positions at which the number of matched positions is generated is divided by the number of matched positions divided by the total number of positions in the comparison window (ie, the window size), and the result is multiplied by 100 to obtain the sequence identity percentage. The term "substantial identity" as used herein represents a characteristic of a polynucleotide or amino acid sequence, wherein the polynucleotide or amino acid comprises a comparison window through at least 18 nucleotide (6 amino acid) positions , Often through a comparison window of at least 24 to 48 nucleotides (8 to 16 amino acids) when compared with a reference sequence with at least 85% sequence identity, preferably at least 90 to 95% sequence identity Sex, more often a sequence with at least 99 sequence identity, where the percent sequence identity is calculated by comparing the reference sequence through a comparison window with sequences that may include deletions or additions (total of 20% or less of the reference sequence) . The reference sequence can be a subset of a larger sequence.

As used herein, the 20 conventional amino acids and their abbreviations follow the conventional usage. See Immunology-A Synthesis (2nd edition, edited by E.S. Golub and D.R. Green, Sinauer Associates, Sunderland 7 Mass. (1991)). Stereo of 20 conventional amino acids, non-natural amino acids (such as α-, α-disubstituted amino acids, N-alkyl amino acids, lactic acid and other non- conventional amino acids) Isomers (such as D-amino acids) may also be suitable components of the polypeptides of the invention. Examples of unconventional amino acids include: 4-hydroxyproline, γ-carboxyglutamic acid, ε-N,N,N-trimethyllysine, ε-N-acetyllysine, O-phosphoserine, N-acetylserine, N-methionine, 3-methylhistidine, 5-hydroxylysine, σ-N-methylarginine And other similar amino acids and imino acids (eg 4-hydroxyproline). In the polypeptide labeling method used herein, the left-hand direction is the amino terminal direction and the right-hand direction is the carboxyl terminal direction in accordance with standard usage and conventions.

Likewise, unless otherwise specified, the left-hand end of a single-stranded polynucleotide sequence is the 5'end, and the left-hand direction of a double-stranded polynucleotide sequence is called the 5'direction. The 5'to 3'joining direction of the nascent RNA transcript is called the transcription direction. The sequence region that has the same sequence as the RNA on the DNA strand and is the 5'pair of the 5'end of the RNA transcript is called the "upstream sequence". The sequence region on the DNA strand that has the same sequence as the RNA and is the 3'end of the 3'pair of RNA transcripts is called the "downstream sequence".

The term "substantial identity" as applied to polypeptides means that two peptide sequences share at least 80% sequence identity when optimally arranged (such as by using the GAP or BESTFIT program with default gap weights). The identity is preferably at least 90% sequence identity, more preferably at least 95% sequence identity, and most preferably at least 99% sequence identity.

The positions of different residues are preferably different due to conservative amino acid substitutions.

Conservative amino acid substitutions refer to the interchangeability of residues with similar side chains. For example, the group of amino acids with aliphatic side chains is glycine, alanine, valine, leucine, and isoleucine; the group of amino acids with aliphatic hydroxyl side chains is serine And threonine; the amino acid groups with side chains containing amide are aspartic acid and glutamic acid; the amino acid groups with aromatic side chains are phenylalanine, tyrosine and tryptamine Acid; Amino acid groups with basic side chains are lysine, arginine and histidine; and amino acid groups with sulfur-containing side chains are cysteine and methionine. The preferred conservative amino acid substitution groups are: valine-leucine-isoleucine, phenylalanine-tyrosine, lysine-arginine, alanine-valine, glutamine Acid-aspartic acid and aspartic acid-glutamic acid.

As discussed herein, it is expected that minor variants in the amino acid sequence of an antibody or immunoglobulin molecule are included in the present invention. The prerequisite is that the amino acid sequence variation is maintained at least 75%, more preferably at least 80%, 90%, 95%, and best 99%. In particular, conservative amino acid substitutions are expected. Conservative substitutions are those that occur within the family of amino acids related to their side chains. The genetically encoded amino acids are usually divided into the following families: (1) Acidic amino acids are aspartic acid and glutamine acid; (2) Basic amino acids are lysine, arginine, and histidine; (3) Non-polar amino acids are alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan; and (4) without electrode amine The base acid is glycine, aspartic acid, glutamic acid, cysteine, serine, threonine, and tyrosine. Hydrophilic amino acids include arginine, aspartic acid, aspartic acid, glutamic acid, glutamic acid, histidine, lysine, serine, and threonine. Hydrophobic amino acids include alanine, cysteine, isoleucine, leucine, methionine, phenylalanine, proline, tryptophan, tyrosine, and valine. Other families of amino acids include (i) serine and threonine, which are aliphatic hydroxyl families; (ii) aspartic acid and glutamic acid, which are amine-containing families; (iii) propylamine Acid, valine, leucine, and isoleucine, which are in the aliphatic family; and (iv) phenylalanine, tryptophan, and tyrosine, which are in the aromatic family. For example, it can be reasonably expected that leucine is replaced by isoleucine or valine alone, aspartic acid is replaced by glutamine alone, threonine is replaced by serine alone, or amino acids are structurally related. Similar substitutions of the amino acids of the amino acid do not have a significant impact on the binding or properties of the resulting molecule, especially if the substitution does not involve the amino acid in the framework site. Whether the amino acid change yields a functional peptide can be easily determined by testing the specific activity of the polypeptide derivative. The verification is detailed in this article. Fragments or analogs of antibodies or immunoglobulin molecules can be easily prepared by those who are generally familiar with the art. The preferred amino and carboxyl ends of the fragments or analogs appear near the boundaries of the functional domains. Structural domains and functional domains can be identified by comparing nucleotide and/or amino acid sequence data with public or private sequence databases. Preferably, computerized comparison methods are used to identify sequence motifs or predicted protein conformation domains that appear in other proteins of known structure and/or function. Methods are known for identifying protein sequences that fold into a known three-dimensional structure. Science 253: 164 (1991) by Bowie et al. Therefore, the foregoing examples prove that those skilled in the art can identify sequence motifs and structural configurations that can be used to define structural domains and functional domains according to the present invention.

Preferred amino acid substitutions are those: (1) reduce the susceptibility to protein degradation, (2) reduce the susceptibility to oxidation, (3) change the binding affinity used to form protein complexes, (4) change Binding affinity, and (5) substitution that confers or modifies the physicochemical or functional properties of these analogs. Analogs may include various muteins of sequences other than the naturally-occurring peptide sequence. For example, single or polyamino acid substitutions (preferably conservative amino acid substitutions) can be achieved in naturally occurring sequences (preferably in the part of the polypeptide outside the domain that forms the intermolecular contact). Conservative amino acid substitutions should not substantially change the structural features of the parental sequence (for example, the replacement of amino acids should not tend to break the helix appearing in the parental sequence, or disrupt other types of secondary sequences that characterize the parental sequence. Level structure). Examples of secondary and tertiary structures of polypeptides recognized by this technology are described in Proteins, Structures and Molecular Principles (edited by Creighton, WH Freeman and Company, New York (1984)); Introduction to Protein Structure (edited by C. Branden and J. Tooze) , Garland Publishing, New York, NY (1991)); and Nature 354: 105 (1991) of Thornton et al.

Peptide analogs are often used in the pharmaceutical industry as non-peptide drugs with properties similar to those of the template peptide. These types of non-peptide compounds are called "peptide mimetic" or "peptidomimetic". Fauchere, J. Adv. Drug Res. 15:29 (1986), Veber and Freidinger TINS p.392 (1985); and Evans et al. J. Med. Chem. 30:1229 (1987). These compounds are often developed with the aid of computerized molecular models. Peptidomimetics that are structurally similar to therapeutically useful peptides can be used to produce equivalent therapeutic or preventive effects. Peptidomimetics are usually structurally similar to paradigm polypeptides (that is, polypeptides with biochemical properties or pharmacological activities), such as human antibodies, but have one or more methods known in the art as needed. A peptide linkage replaced by a linkage selected from the group consisting of: --CH ₂ NH--, --CH ₂ S-, --CH ₂ -CH ₂ --, --CH=CH- -(Cis and trans), --COCH ₂ --, CH(OH)CH ₂ - and -CH ₂ SO--. Systemic substitution of one or more amino acids of the same sequence with the same type of D-amino acid (for example, replacement of L-lysine with D-lysine) can be used to produce more stable peptides. In addition, constrained peptides containing consensus sequences or substantially identical consensus sequence variations can be produced by methods known in the art (Rizo and Gierasch Ann. Rev. Biochem. 61:387 (1992)); for example, by By adding internal cysteine residues that can form intramolecular disulfide bridges that cyclize the peptide.

The term "agent" as used herein refers to a chemical compound, a mixture of chemical compounds, a biological macromolecule, or an extract made from biological materials.

The term "label" or "labeled" as used herein refers to the incorporation of a detectable marker, such as the incorporation of a radiolabeled amino acid or the attachment of biotin that can be detected by labeled avidin Part of the polypeptide (for example, streptavidin containing fluorescent markers or enzymatic activity that can be detected optically or calorimetrically). Under certain circumstances, the marker or marker can also be therapeutic. Various methods of labeling polypeptides and glycoproteins are known in the art and can be used. Examples of labels for polypeptides include, but are not limited to, the following: radioisotopes or radionuclides (e.g. ³ H, ¹⁴ C, ¹⁵ N, ³⁵ S, ⁹⁰ Y, ⁹⁹ Tc, ¹¹¹ In, ¹²⁵ I, ¹³¹ I) , Fluorescent markers (such as FITC, rose bengal, lanthanum element phosphors), enzyme markers (such as wasabi peroxidase, p-galactosidase, luciferase, alkaline phosphatase), chemiluminescence, Biotin group, a predetermined polypeptide epitope recognized by a secondary reporter (such as leucine zipper pair sequence, secondary antibody binding site, metal binding domain, epitope tag). In some embodiments, the markers are connected by spacer arms of various lengths to reduce potential steric hindrance. The term "medical agent or drug" as used herein refers to a chemical compound or composition that can induce the desired therapeutic effect when properly administered to a patient.

Other chemical terms herein are used according to the conventional usage of the technology, as exemplified by The McGraw-Hill Dictionary of Chemical Terms (Parker, S., Ed., McGraw-Hill, San Francisco (1985)).

As used herein, "substantially pure" means that the target species is the species that mainly exists (that is, it is more abundant on a molar basis than any other individual species in the composition), and the substantially purified part is better It is a composition in which the target species accounts for at least about 50% (on a molar basis) of all existing macromolecular species.

The substantially pure composition generally accounts for more than about 80% of all macromolecular species present in the composition, more preferably about 85%, 90%, 95%, and 99%. The target species is better purified to the necessary homogeneity (contaminant species cannot be detected in the composition by conventional detection methods), and the composition is basically composed of a single macromolecular species.

The articles "a", "an" and "the" used in both the following description and the scope of the patent application are interpreted as covering both the singular and the plural, unless otherwise indicated herein or clearly negated by the context. The terms "including", "having", "being of", "including" and "containing" in "has a chemical formula" are also interpreted as open-ended terms (that is, meaning "including but not Limited to "), unless otherwise noted. For example, a polymer scaffold of a specific formula includes all monomer units shown in the formula and may also include additional monomer units not shown in the formula. In addition, whenever "comprising" or another open-ended term is used in the implementation, it should be understood that the intermediate term "essentially composed of" or the closed term "consisted of" can be used "More rigorously advocate the same implementation style.

When the terms "about", "approximately" or "approximate" are used in conjunction with numerical values, these terms are meant to include numerical sets or numerical ranges. For example, "about X" includes ±20%, ±10%, ±5%, ±2%, ±1%, ±0.5%, ±0.2%, or ±0.1% of X, where X is a numerical value. In one embodiment, the term "about" refers to a numerical range that is 5% more or less than the specified value. In another embodiment, the term "about" refers to a numerical range that is 2% more or less than the specified value. In another embodiment, the term "about" refers to a numerical range that is 1% more or less than the specified value.

The recitation of a range of values is only intended to serve as a shorthand method for each individual value to be individually recited within the range, unless otherwise indicated herein, and each individual value is incorporated into the specification as if it were individually recited herein . The ranges used herein include both ends of the range unless otherwise specified. For example, the phrases "x is an integer between 1 and 6" and "x is an integer from 1 to 6" both mean "x is 1, 2, 3, 4, 5, or 6", that is, the term " "Between X and Y" and "X to Y range" include X and Y and integers in between.

All methods described herein can be performed in any suitable order, unless otherwise indicated herein or clearly denied by context. The use of any and all examples or illustrative language (such as "such as") provided herein is only intended to better exemplify the present invention and should not be construed as a limitation on the scope of the patent application, unless otherwise expressly claimed. The language in the specification should not be interpreted as indicating that any unclaimed element is necessary to be claimed.

"Polymer carrier or stent": as used herein, the term polymer carrier or stent refers to a drug molecule suitable for covalent attachment or covalent attachment of one or more drug molecules with labeled linkers and/or one or more A polymer or modified polymer of PBRM with the indicated linker.

"Physiological conditions": As used herein, the phrase "physiological conditions" refers to the range of chemical (for example, pH, ionic strength) and biochemical (for example, enzyme concentration) conditions that may be encountered by the extracellular fluid of living tissues. For most normal tissues, the physiological pH is in the range of about 7.0 to 7.4. Circulating plasma and normal interstitial fluid represent typical examples of normal physiological conditions.

"Drug": The term "drug" as used herein refers to a compound (such as an active pharmaceutical ingredient) that has biological activity and provides a desired physiological effect after administration to an individual in need thereof.

"Cytotoxicity": The term "cytotoxicity" as used herein means toxicity to cells or test cell populations (eg cancer cells). Toxic effects can cause cell death and/or lysis. In certain cases, the toxic effect may be a sublethal destructive effect on the cell, such as slowing down or stopping cell growth. In order to achieve the cytotoxic effect, the drug or prodrug can be selected from the group consisting of DNA damaging agents, microtubule disrupting agents, or cytotoxic proteins or polypeptides.

"PHF" refers to poly(1-hydroxymethylethylene hydroxymethyl-formal).

As used herein, the terms "polymer unit", "monomeric unit", "monomer", "monomer unit" and "unit" all refer to the repeatable structure of the polymer unit.

The "molecular weight" or "MW" of the polymer or polymer carrier/stent or polymer conjugate as used herein refers to the weight average molecular weight of the unmodified polymer, unless otherwise specified.

As used herein, "dosing regimen" or "dosage regimen" refers to the amount of agent administered (for example, a composition containing a polymer antibody-drug conjugate targeting NaPi2b) and the frequency of administration. The dosage regimen is a function of the disease or condition to be treated and can therefore be changed.

The "frequency" of administration as used herein refers to the time between consecutive administrations of treatment. For example, the frequency can be days, weeks, or months. For example, the frequency can be more than once a week, such as twice a week, three times a week, four times a week, five times a week, six times a week, or every day. The frequency can also be one, two, three or four weeks. The specific frequency is a function of the specific disease or condition being treated. The frequency is usually more than once a week and usually twice a week.

The "administration cycle" as used herein refers to the repeated time course of a dosing regimen of repeated continuous administration of the enzyme and/or the second dose. For example, an exemplary administration cycle is a 28-day cycle in which administration is twice a week for three weeks, and then administration is stopped for a week. The preferred dosage cycle is a 21-day cycle of once every 21 days (ie, 3 weeks) or a 28-day cycle of once every 28 days (ie, 4 weeks).

As used herein, when referring to an individual-based dose of mg/kg, the average human individual is considered to have a mass of about 70 kg to 75 kg (such as 70 kg) and a body surface area (BSA) of 1.73 m ² .

As used herein, ameliorating the symptoms of a particular disease or condition by treatment (such as administration of a pharmaceutical composition or other therapeutic agent) refers to any alleviation (whether permanent or temporary, continuous or transient) of the symptoms or adverse effects of the condition, Such as reducing the adverse effects associated with or during the administration of the polymer antibody-drug conjugate targeting NaPi2b.

As used herein, when referring to a dose based on "body surface area"(BSA; m ² ), it is the measured or calculated surface area of the human body. For many clinical purposes, BSA is a better metabolic quality indicator than body weight because it is less affected by abnormal fat mass. A variety of calculation methods for obtaining BSA have been published without direct measurement. In the following formula, BSA is measured in m ² , W is the mass in kg, and H is the height in cm. The most widely used Du Bois, Du Bois formula: BSA = 0.007184 x W ^0.425 x H ^0.725 . Other methods for determining BSA include, for example, Mosteller, Haycock, Gehan and George, Boyd, Fujimoto, Takahira, Shuter and Aslani or Schlich formula.

As used herein, "treating" or "treat" refers to the management and care of patients for the purpose of combating diseases, conditions or disorders, and includes administration of the conjugate of the present invention or its combination with Immunomodulatory treatments (e.g., immuno-oncology agents, such as immune checkpoint inhibitors) are combined with pharmaceutical compositions to relieve symptoms or complications of diseases, conditions, or disorders, or to remove diseases, conditions, or disorders.

As used herein, "prevention" or "prophylaxis" refers to reducing the risk of developing a disease or condition, or reducing or removing the onset of symptoms or complications of a disease, condition or condition.

When the term "effective amount" or "sufficient amount" refers to the active agent, the term refers to the amount necessary for the desired biological response. As used herein, "therapeutically effective dose" or "therapeutically effective dose" refers to the amount or quantity of an agent, compound, material, or composition containing the compound that is at least sufficient to produce a detectable therapeutic effect. The effect can be detected by any test known in the art. The precise effective amount for an individual depends on the individual's weight, body type, and health; the nature and extent of the condition; and the therapeutic agent selected for administration.

"Individual" includes mammals. The mammal can be, for example, any mammal (e.g., human), primate, bird, mouse, rat, poultry, dog, cat, cow, horse, goat, camel, sheep, or pig. The mammal is preferably a human.

As used herein, "unit dose form" or "unit dosage form" refers to physically discrete units suitable for individual humans and animals and individually packaged, as known in the art.

A single dose formulation as used herein refers to a formulation that is a single dose.

As used herein, “temporal proximity” means that the administration of a therapeutic agent (such as the polymer antibody-drug conjugate targeting NaPi2b disclosed herein) occurs in another therapeutic agent (such as In the period before or after the administration of the immune checkpoint inhibitors disclosed herein, the therapeutic effect of one therapeutic agent overlaps with the therapeutic effect of another therapeutic agent. In some embodiments, the therapeutic effect of one therapeutic agent completely overlaps with the therapeutic effect of another therapeutic agent. In some embodiments, "sequential proximity" means that the administration of one therapeutic agent occurs within a time period before or after the administration of another therapeutic agent, so that the therapeutic effect of one therapeutic agent and the other therapeutic agent There is synergy between. "Sequential proximity" can be changed according to a variety of factors, including but not limited to the age, gender, weight, genetic background, medical condition, disease history and treatment history of the individual to be administered the therapeutic agent; the disease or condition to be treated or improved ; The therapeutic outcome to be achieved; the dosage, frequency and duration of administration of the therapeutic agent; the pharmacokinetics and pharmacodynamics of the therapeutic agent; and the route of administration of the therapeutic agent. In some embodiments, "timing proximity" means within 15 minutes, within 30 minutes, within 1 hour, within 2 hours, within 4 hours, within 6 hours, within 8 hours, Within 12 hours, within 18 hours, within 24 hours, within 36 hours, within 2 days, within 3 days, within 4 days, within 5 days, within 6 days, within 1 week, within Within 2 weeks, within 3 weeks, within 4 weeks, within 6 weeks, or within 8 weeks. In some embodiments, multiple administrations of one therapeutic agent may occur close to the single administration of another therapeutic agent in time sequence. In some embodiments, the timing proximity can be changed during the treatment cycle or within the dosing schedule.

As used herein, "kit" refers to a combination of components, such as the composition herein for the purpose of including but not limited to reconstitution, activation and for delivery, administration, diagnosis, and assessment of biological activity or properties The purpose of the appliance/device in combination with another item. The kit includes instructions as needed.

The present invention is intended to include all isotopes of atoms present in the compounds of the present invention. Isotopes include those atoms that have the same atomic number but different mass numbers. By way of general example and non-limiting illustration, isotopes of hydrogen include tritium and deuterium. Isotopes of carbon include C-13 and C-14.

The present invention is intended to include all isomers of compounds, which refer to and include optical isomers and tautomers, wherein optical isomers include enantiomers and diastereomers, chiral isomers and non-enantiomers. Chiral isomers, and optical isomers include single-isolated optical isomers and mixtures of optical isomers, including racemic and non-racemic mixtures; wherein the isomers can be in single-isolated form or with A mixture of one or more other isomers. Other implementation styles

All publications and patent documents cited herein are incorporated herein for reference, as if each such publication or document was specifically or individually indicated and incorporated herein for reference. The quotation of publications and patent documents is not intended to recognize any of them as relevant prior art, nor does it constitute any recognition of their content or date. The present invention has been described in written descriptions. Those familiar with the technical field should recognize that the present invention can be implemented in various embodiments and the foregoing description and the following examples are for the purpose of exemplifying and not limiting the scope of subsequent patent applications. Example

The following examples are illustrative and not intended to be limiting, and those skilled in the art can easily understand that other reagents or methods can be used. Abbreviations

The following abbreviations are used in the subsequent reaction schemes and synthesis examples. This list does not represent all the abbreviations used in the application are included in the list, because those familiar with the field of organic synthesis technology can easily understand that additional standard abbreviations can also be used in the synthesis process and examples. AF-HPA Auristatin F-Hydroxypropylamide BSA Body surface area CR Complete response DCR Disease control rate DES Dose escalation DLT Dose limiting toxicity EXP Team expansion IHC immunochemistry IV Intravenous MTD Maximum tolerated dose NSCLC Non-small cell lung cancer ORR Objective response rate PBS Phosphate buffered saline PE Body checkup PHF Poly(1-hydroxymethyl ethylene hydroxymethyl formal) PR Partial response RP2D Recommended dose for Phase 2 SD Stable disease SRC Security Review Committee SRM Security Review Meeting TRAE Treatment-related adverse events General Information

XMT-1536 is prepared as described in US Application No. 2017/0266311.

AF-HPA is prepared as described in US Patent No. 8808679 (B2).

CDR is identified in Kabat numbering format. Example 1: Evaluation of safety and tolerance of XMT-1536 Research design

The study presented in this article is an open-label, multicenter Phase Ib study of XMT-1536 administered by intravenous infusion every three or four weeks. The dose escalation (DES) part of the study is to establish the maximum tolerated dose (MTD) or phase 2 recommended dose (RP2D) of XMT-1536 in patients with several tumor types that may show NaPi2b, focusing on patients with platinum Patients with drug-resistant ovarian cancer and non-squamous non-small cell lung cancer (NSCLC). MTD is defined as the highest dose of XMT-1536 that does not cause unacceptable toxicity defined by the planned dose-limiting toxicity guidelines. RP2D may be different from MTD and may be based on RECIST 1.1 to consider the tolerability, pharmacokinetics and pharmacodynamic measurements and efficacy of XMT-1536. After the first cycle, if the researcher believes that the drug is well tolerated and the patient continues to obtain clinical benefits, the patient can continue to receive XMT-1536 until the disease progresses. The DES stage of the study uses an accelerated titration design. The first 3-week cycle of treatment constitutes the dose-limiting toxicity (DLT) evaluation period. Regarding the first two dose levels, at least 1 patient is treated at each dose level. If this patient experiences an adverse event after undergoing grade 2 or higher treatment, two additional patients will be added at this dose level and the follow-up of the study will follow the 3+3 design, as described below. If the patient has not experienced a treatment-related event or DLT of grade 2 or higher during the DLT evaluation period and the Safety Review Committee (SRC) agrees that this is a reasonable well-tolerated dose, then it will be selected at the next dose level. Starting with dose level 3, the study will follow the standard 3+3 design, initially enrolling 3 patients at each dose level. If none of the 3 patients experienced DLT during the evaluation period and the SRC agreed that this was a reasonable well tolerated dose, then 3 patients were enrolled at the next dose level. However, if 1 DLT occurs, 3 other patients will be enrolled at the same dose level. Any dose level with 2 or more DLT is considered to exceed the MTD, and subsequent patients will be selected at the lower dose level. After the first cycle, if the researcher believes that the drug is well tolerated and the patient continues to obtain clinical benefits, the patient can continue to receive XMT-1536 until the disease progresses.

After completing DES, the EXP stage of the study will be opened to the three groups of patients. Team 1 accommodates patients with platinum drug-resistant ovarian cancer. Team 2 accommodates patients with non-squamous NSCLC. Team 3 accommodates patients with papillary thyroid cancer, endometrial cancer, papillary renal cell carcinoma, or salivary duct cancer.

After the planning revision in August 2018, each cycle is a 4-week (28-day) dose cycle. The first two 4-week cycles (56 days) of treatment constitute a dose-limiting toxicity (DLT) evaluation period. Starting with this revision, up to 10 patients were selected for the new 4-week cycle dosing regimen evaluated at 20.0 mg/m ² . If the dose is approved by the SRC, the subsequent dose groups will follow the revised 3+3 design. Three patients were initially enrolled, and if toxicity issues arise, the SRC will review all relevant data and may decide to add more patients to fully evaluate the dose under discussion and/or lower doses. If the dose is well tolerated and 3 patients come to the end of cycle 2 without DLT, the SRC may approve this dose and open the next dose. The observation period of DLT is 56 days between the first day and the end of the second cycle, which includes the pre-dose assessment before receiving the third cycle dose. After at least 6 patients administered at each dose level complete the second cycle, and no later than 5 days after the 28-day evaluation before SRM, a maximum of 10 patients will be selected for each dose level. The effective data of all previously administered patients are included in the data review of the dose-escalation SRM of each application. Any dose level of 2 or more DLT in 6 or fewer treated patients is considered to exceed the MTD, and subsequent patients will be selected at the lower dose level. After the second cycle, if the researcher believes that the drug is well tolerated and the patient continues to obtain clinical benefits, the patient can continue to receive XMT-1536 until the disease progresses. After the completion of DES, the EXP stage of open research.

All adverse events were graded according to the National Cancer Institute (NCI) Common Terminology Guidelines Version (CTCAE v4.03). The observation period of DLT is 56 days between the first day and the end of the second cycle, which includes the pre-dose assessment before receiving the third cycle dose. Generally speaking, adverse event grade ≥ 3 is a DLT modified to the following criteria: neutropenia blood toxicity, gastrointestinal toxicity, liver toxicity and electrolyte imbalance. XMT-1536-related toxicity that delays the initiation of cycle 3 for more than 2 weeks, hospitalization to treat infusion-related reactions, and any toxicity that prompts the modification of the dose to be administered in cycle 3 is also DLT. Blood samples were taken to determine the plasma PK parameters of XMT-1536, its release product Auristatin F-HPA, and selected metabolites. Perform anti-drug antibody (ADA) test and neutralizing antibody (nAb). The tumor response was evaluated by the researcher at the end of the second cycle and every 2 subsequent cycles using the Response Evaluation Criteria for Solid Tumors (RECIST) version 1.1. Research visit

A research visit will be conducted. If the safety assessment result at the end of the cycle is obtained and reviewed before the start of the next cycle, the last day of the current cycle can be the same day as the beginning of the next cycle of the entire study. Researchers can arrange accommodation during the visit based on the patient's medical needs. Before making any adjustments, discuss possible schedule changes with the medical monitor, unless it is an urgent need. The researchers evaluated each patient before initiating the XMT-1536 dose, as described in the evaluation schedule. A complete physical examination (PE) consists of examining the head/ears/eyes/nose/throat (HEENT), palpable tumors, nervous and muscular systems, lungs and cardiovascular systems, abdomen and lower limbs. A brief physical examination consists of examinations: HEENT, lungs and cardiovascular system. Evaluate the patient's condition or new and/or worsening toxicity at each visit. All adverse events were graded according to the National Cancer Institute (NCI) Common Terminology Guidelines Version (CTCAE v4.03). Number of individuals

The exact number of patients to be selected for DES cannot be predicted and depends on when MTD or RP2D is reached. Based on preclinical animal studies, it is reasonably expected that 21-40 patients will be administered DES. The EXP stage of the study plans to treat up to 30 patients in each of the 3 disease teams. Given the dropout rate, it is reasonable to expect that 90 to 105 patients will be administered EXP. qualifications

Patients selected as research candidates are evaluated by the researcher to ensure that they meet the inclusion and exclusion criteria and that the patients are eligible to participate. Table 1. Inclusion criteria for DES and EXP Numbering Eligibility criteria 1 Female and male, age ≥18 years old. 2 ECOG performance status 0 or 1. 3 Measurable diseases according to RECIST, version 1.1. 4 All acute toxic effects of previous treatment or surgery subsided to grade ≤ 1 (except for hair removal). 5 The left ventricular ejection rate (LVEF) of the heart is ≥50% or the lower limit of normal of the organization obtained by Echo or MUGA scan. 6 Appropriate organ function as defined by the following criteria: absolute neutrophil count (ANC) ≥ 1500 cells/mm ³ , platelet count ≥ 100,000 cells/mm ³ , heme ≥ 9 g/dL, INR, activated fraction Thrombin time (aPTT) and prothrombin time (PT) are all within the upper limit of normal (ULN) of the institution, serum creatinine ≤1.5 mg/dL or calculated creatinine clearance rate ≥60 mL/min ¹ , total bile Red pigment ≤ ULN, aspartate aminotransferase (AST or SGOT) and alanine aminotransferase (ALT or SGPT) ≤ 1.5 times the upper limit of normal (ULN). Albumin ≥3.0 g/dL. ¹ Calculated by the Cockroft and Gault method. Creatinine clearance rate (mL/min) = (140-age) x body weight (kg)/72x (serum creatinine in mg/dL) = mL/min (for women, multiply the result by 0.85). 7 Tumor tissue block (strongly recommended) or freshly sliced tissue section (refer to the laboratory manual) confirmed usability in NaPi2b test and laboratory assessment (in cycle 1, before day 1). The tissue sample must be submitted within 45 days after the first dose of the study drug. 8 For women with fertility and men with fertile partners, the patient and/or partner agree to use a highly effective form of hormonal contraceptives or two effective forms of non-hormonal contraceptives, and during the study treatment period and at the end of the study treatment Continue to use birth control pills for at least 6 months after the dose. Male patients whose partners are pregnant should use condoms during pregnancy. 9 Able to sign informed consent.

Preventive blood transfusion (or blood components) before the initial administration should not be used to meet the selection criteria. Blood transfusion (or blood components) is allowed to control and treat sudden anemia or other cytopenias and should be recorded as a concomitant medication. Do not use growth factor prophylactics before XMT-1536 is administered in any cycle. Table 2: Disease-specific inclusion criteria for DES Numbering Eligibility criteria 1 The following types of solid tumors confirmed by histology or cytology for incurable, locally advanced, or metastatic disease that have failed standard treatment or have no standard treatment options: Platinum drug-resistant ovarian cancer (including epithelial ovarian cancer, such as high-grade serous Ovarian cancer, fallopian tube and primary peritoneal cancer), non-squamous NSCLC, papillary thyroid cancer, endometrial cancer (excluding carcinosarcoma and stromal tumor), papillary renal cell carcinoma, salivary gland duct cancer. Table 3: Disease-specific inclusion criteria for the expansion stage (EXP) Numbering Eligibility criteria Team 1: Platinum drug-resistant ovarian cancer 1 Histological diagnosis of high-grade serous ovarian, epithelial ovarian, fallopian tube or primary peritoneal cancer, excluding mucinous subtypes. 2 Platinum drug resistance is defined as disease progression within 6 months of completing platinum-containing chemotherapy. 3 No more than 3 lines of previous treatment Team 2: Non-squamous NSCLC 1 Histological diagnosis of adenocarcinoma of non-squamous NSCLC 2 Previous treatment (combined or continuous) with platinum-based (cisplatin or carboplatin) regimen and PD-1 or PD-L1 monoclonal antibody. 3 Patients with known cancer-causing mutations (such as ALK translocation, EGFR mutations) for approved treatments must prove intolerance or disease progression to the approved treatments for their mutations. Patients must receive prior treatment with platinum-based regimens, but do not require prior treatment with PD-1 or PD-L1 monoclonal antibodies. 4 No previous treatment with cytotoxic agents or immunotherapy. Team 3: Additional indications 1 For patients with papillary thyroid cancer, the following should be met: (a) progressive, radioactive iodine refractory, locally recurrent or metastatic disease, and (b) treatment of previous kinase inhibitors (such as Lavatinib ( Lenvatinib) and sorafenib (sorafenib) are resistant or intolerant. Patients deemed unsuitable for kinase inhibitor therapy can be selected upon approval by a medical monitor. 2 For patients with endometrial cancer, the following are required: (a) The diagnosis of epithelial endometrial cancer is required. Stromal tumors and carcinosarcomas (Mullerian mixed tumors) are excluded, and (b) patients must receive at least one previous chemotherapy regimen of carboplatin/paclitaxel or similar regimens for endometrial cancer. If appropriate, patients should receive prior hormone therapy for endometrial cancer, such as low-grade, hormone receptor-positive endometrial adenocarcinoma. 3 For patients with papillary renal cell carcinoma, the following should be met: (a) local confirmation of renal cell carcinoma with a significant papillary growth pattern, and (b) progression after standard systemic treatment. 4 Regarding patients with salivary duct cancer, the following should be met: (a) histological diagnosis of salivary duct cancer (excluding other subtypes of salivary gland cancer), and (b) the researcher's assessment of progress or lack of available after standard systemic treatment Effective treatment. Table 4: Exclusion criteria for dose escalation and amplification Numbering Eligibility criteria Exclusion criteria for dose escalation and amplification 1 Major surgery within 28 days of starting study treatment; or prior treatment of 28 days or 5 half-lives before starting study treatment (14 days or 5 half-lives for small molecule targeted therapy), whichever is shorter Anti-cancer therapy; or recent radiotherapy with unresolved toxicity or within a time window of potential toxicity (it is recommended to consult with the medical monitor). 2 Brain metastases: untreated, progressive or require any type of primary treatment, such as whole brain radiation therapy, adjuvant chemotherapy, gamma knife to control symptoms from brain metastases within 30 days of the initial study treatment, or Any history of meningeal metastases. 3 Active infection with HIV, hepatitis B virus (HBV) or hepatitis C virus (HCV) currently known. In addition, negative serological tests are required during the screening of HBV and HCV: a. HBV: The patient's hepatitis B surface antigen (HBsAg) and hepatitis B nuclear antibody (anti-HBc) must be negative. Patients with evidence of previous HBV infection (positive anti-HBc and negative HBsAg) must have negative HBV DNA to be considered eligible. b. HCV: The patient's HCV antibody must be negative, or if the HCV antibody is positive, the patient must undergo a negative PCR test of HCV RNA. 4 Current serious, uncontrolled systemic diseases (such as clinically significant cardiovascular, pulmonary or metabolic diseases) or intermittent diseases may interfere with planned assessments. 5 History of liver cirrhosis, liver fibrosis, varicose veins, or other clinically significant liver diseases. Patients with a history of chronic liver disease (such as fatty liver) may require fiber scan testing. Women with a fixed daily alcohol intake of more than one cup or men with more than two cups today are not eligible and should not be encouraged to drink during the trial period. 6 Patients cannot receive drugs associated with liver toxicity at the same time as XMT-1536 is administered or within 21 days after the last dose of XMT-1536. Patients can receive acetaminophen/paracetamol for a limited time, but the total daily dose is ≤2 g per day. Encourage the use of NSAIDs or steroids to treat fever. 7 Severe breathing difficulties or need for supplemental oxygen therapy at rest due to complications of advanced malignancies. 8 Current active pneumonia or interstitial lung disease. 9 Pregnant or breastfeeding women. 10 Diagnosis of secondary malignant tumors that may require systemic anticancer therapy. 11 Active corneal disease or conjunctival disease or a history of corneal disease or conjunctival disease within 12 months prior to enrollment. 12 Use potent CYP450 inhibitors. Exclusion criteria for amplification 10 Use potent CYP450 inhibitors. Drug use associated with liver toxicity

FDA-classified drugs that are most likely to cause drug-induced liver injury (DILI) should not be used during the study period. The exception to the use of paracetamol (acetaminophen) is described in exclusion criterion 6. If a drug on the FDA list must be used during the study to give proper medical care and no alternative is available, the medical monitor must discuss the situation. Individual withdrawal criteria

Patients are free to withdraw from the study at any time.

If possible, patients who want to terminate study participation early are required to undergo general medical follow-up at least 30 days after administration for safety monitoring purposes. This requirement is prioritized for entry into tranquility care. Follow the procedures described for the end of the treatment visit.

In all cases, the reason for withdrawal must be recorded in the eCRF. If the cause is not known initially, the patient should be followed to establish whether the cause is an adverse event. If so, this event was recorded as the reason for the termination of the study. Dosage and administration

XMT-1536 is a colorless to yellow or brown liquid provided in a 5 mL round flint glass vial. The vial has a gray chlorobutyl rubber stopper with a 20mm green aluminum flip cap covering the barrier. Each single-use vial contains 2.5 mL of XMT-1536 antibody drug conjugate at a concentration of 10 mg/mL and a pH of 4.0 to 6.0.

The vials must be stored in a safe, temperature-controlled freezer at -20°C (±5°C). XMT-1536 vials are the preparation and control of standard pharmaceutical procedures for institutions that use controlled research substances. The assigned CRA is to review storage conditions and maintenance of these conditions during regular site visits.

Before preparing the patient's dose, each individual vial of XMT-1536 was allowed to thaw at room temperature for up to 60 minutes. Check the thawed vial before use: the liquid should be transparent to yellow to brown, with almost no visible particles. Do not shake the vial or place it in direct sunlight. Once the infusion dose is prepared, it can be left at room temperature under typical indoor lighting conditions for up to 4 hours before administration.

XMT-1536 is administered based on body surface area (BSA). The dose adjusted by BSA is calculated in accordance with the standard practice of each agency. When possible, use the Mosteller formula. The starting dose is calculated based on the height and weight collected within 14 days of the first dose (can be collected on the day of the first dose). Dosage adjustments based on subsequent body weight measurements are carried out in accordance with the standard practices of various institutions. If there is no standard practice, gain additional body weight and confirm or change BSA before the second cycle and every 2 cycles thereafter.

Each dose is prepared in a 100 mL, 0.9% NS PVC infusion bag. Dose preparation is performed according to the procedures at the study site. Refer to the Pharmacy Manual for detailed instructions. Dose preparation is recorded in each patient’s study participation record and follows all institutional practices, including quality assurance checks.

The planned starting dose in the dose escalation part of the study is 3 mg/m ² XMT-1536 ADC. The dose escalation procedure is shown in Table 5 below. As of August 2018, DL 5 (30 mg/m ² ) has been approved by SRC. However, if the dose is found to be tolerable, the SRC decided to extend the DLT observation period to a two-week period (56 days) based on the newly emerging safety data, revise the inclusion and exclusion criteria, and add additional patients Under DL 4 (20 mg/m ² ), the purpose is to restart the dose escalation in accordance with the planning guidelines. SRC can decide: (1) Increase the dose to the next planned level or a higher dose level that is less than the planned increase based on the data that proves the drug tolerability. (2) Add up to 3 patients to the current dose level for additional evaluation. (3) Decrease to a lower dose level for further evaluation; the previously agreed dose level or a new lower dose level can be selected based on the safety data of the patient. (4) MTD has been reached and no further dose escalation has occurred. Before starting the expansion phase of the study, at least 6 patients were treated with MTD or RP2D. The SRC may be determined to be increased in smaller increments than shown in Table 5 after reviewing all relevant patient data at any or all dose levels depending on the tolerance of the given dose level. Table 5. Dose escalation levels Dose escalation Start 100% 100% 67% 50% 20% 20% Dose level (DL) 1 2 3 4 4A 5 5A 6 6A 7A Initial number of patients 1 1 3 3 3 3 3 Planned dose, mg/m ² 3 6 12 20 30 40(DL 6) 36(DL 6A) 43

The dose adjusted by BSA is calculated in accordance with the standard practice of each agency. When possible, use Mostler's formula. The starting dose is collected within 14 days of the first dose and can be calculated based on the height and weight collected on the day of the first dose. Dosage adjustments based on subsequent body weight measurements are carried out in accordance with the standard practices of various institutions. If there is no standard practice, gain additional body weight and confirm or change BSA before the second cycle and every 2 cycles thereafter.

Conventional supportive care medications can be used to treat dose-related concurrent acute conditions, such as allergic reactions, vomiting, diarrhea, and fever. It is not recommended to perform preventive treatment of infusion reactions or allergies before the first dose of any patient, but if these reactions occur after the first dose, the preventive treatment can be administered before the second dose and/or subsequent doses treatment. Treatment with antiemetics and antipyretics can be necessary within a few days after infusion. If the patient's medical history shows that it is necessary, it can be used before the first dose. Under certain circumstances, it is allowed to adjust the dose after the first cycle.

The initial dose for each patient was administered within 90 minutes. If no infusion-related reaction (IRR) occurs, all subsequent doses will be administered within 30 to 90 minutes at the discretion of the researcher. The treatment of IRR should be based on institutional standards or research guidelines.

Once the patient has completed the first cycle (DLT evaluation period), if the patient experiences toxicity, but the researcher believes that the patient will benefit from further treatment with XMT-1536, the dose can be adjusted. Guidelines for continuous administration

In cycle 2 and beyond, the researcher or designer should review the clinical and laboratory findings at the end of the cycle/pre-dose before continuing the study drug administration. All liver enzymes should be restored to grade 1 or lower or to the patient's baseline before proceeding. Laboratory data (especially liver tests) should be compared with the patient's baseline, and a delay in dosing (e.g. 1 week) should be considered to allow more complete recovery of liver function or to allow other AEs to resolve, such as nausea, vomiting, anorexia, or fatigue. Reduce the dose of the same patient (Intrapatient)

Once the patient has completed the first cycle, if the patient experiences toxicity, but the researcher believes that the patient will benefit from further treatment with XMT-1536, the dose can be adjusted. Only when the observed toxicity resolves to Gr≤1 or baseline within 21 days or 28 days after the onset of toxicity, and after discussion with the researcher and the medical monitor, as appropriate, treatment can be initiated at a reduced dose (Table 6).

The first dose reduction was one dose lower than the dose level at which toxicity was observed.

The second reduction was two doses lower than the original dose at which toxicity was observed.

If a third dose reduction is required, the patient is stopped. Table 6. Dose reduction after completing cycle 1 Reduce the dose Dose ¹ 1 ^st One dose level lower than the original dose 2 ^nd Two dose levels lower than the original dose 3 ^rd Patient stopped ^1The reduced dose should not be less than 3 mg/m ² (DL 1)

In the second cycle or subsequent cycles, peripheral motor or sensory neuropathy grade ≥ 3 that does not resolve to grade ≤ 2 within 21 days after the situation requires dose reduction or termination of treatment. The medical monitor should usually be called to discuss before making any dose adjustments.

In cycle 2 and beyond, the researcher or designer should review the clinical and laboratory findings at the end of the cycle/pre-dose before continuing the study drug administration. All liver enzymes should be restored to grade 1 or lower or to the patient's baseline before proceeding. Laboratory data (especially liver tests) should be compared with the patient's baseline, and a delay in dosing (e.g. 1 week) should be considered to allow more complete recovery of liver function or to allow other AEs to resolve, such as nausea, vomiting, anorexia, or fatigue. Reduce the dose in DES and EXP due to liver toxicity

Patients who have experienced hepatic transaminase or ALP levels of grade ≥3 in both DES and EXP will return to the office, retest within 48 to 72 hours of the initial grade value ≥3 and monitor at least weekly until the content returns to grade ≤1 or reach patient baseline. If the patient lives in a place far away from the research facility, making it difficult to return, he can arrange for a blood draw locally, and return the results and local normal ranges to the researcher for monitoring.

Patients who have experienced hepatic transaminase or ALP increase to grade ≥ 3 and need more than 72 hours to recover to grade ≤ 2 (or baseline) can continue to be administered in the next cycle, but at the next lower dose.

If the patient experiences a second hepatic transaminase or ALP increase to grade ≥ 3 after a previous dose reduction, and it takes more than 72 hours to recover to grade ≤ 2 (or baseline), he or she can be in the next cycle, but The administration is continued at the next dose lower than the dose when the second increase occurred.

If the patient experiences a third hepatic transaminase or ALP increase to grade ≥3 after the previous reduction of two doses, and it takes more than 72 hours to recover to grade ≤2 (or baseline), he or she will stop the study and Monitor at least weekly until the content returns to level ≤ 1 (or baseline).

If it takes less than 72 hours to recover to grade ≤ 2 (or baseline), the XMT-1536 dose will not be modified due to liver transaminase or ALP increase to grade ≥ 3.

Any patient experiencing Gr 4 liver transaminase or ALP will stop the study and will not receive dose reduction. Monitor Gr 4 toxicity at least weekly until return to Gr≤1 or baseline.

Regardless of whether the threshold for dose reduction is reached, any hepatic transaminase or ALP that increases beyond grade 1 must return to grade ≤ 1 or baseline before the next study drug dose. Duration of treatment

Treatment will continue indefinitely, unless one of the following occurs: disease progression, intermittent disease that prevents further administration of treatment, unacceptable adverse events or pregnancy, patient non-compliance with the plan, patient withdrawal consent to participate in the study and/or overall or Certain patient condition changes make the patient unacceptable for further treatment under the judgment of the researcher.

However, if disease progression occurs, but the researcher realizes that the patient may benefit from continuous exposure to XMT-1536, the researcher may choose to continue the administration after discussing with the medical monitor. main target

The main goal of the dose escalation (DES) study is to determine the maximum tolerated dose (MTD) and phase 2 recommended dose (RP2D) of XMT-1536 administered intravenously every three weeks or every four weeks and assess the safety of XMT-1536 Sex and tolerance.

The main goal of the expansion study is to further evaluate the safety and tolerability of XMT-1536 administered with MTD/RP2D identified by DES, and to evaluate the preliminary anti-neoplastic activity of XMT-1536. Secondary goal

The secondary goal of the dose escalation (DES) study is to assess the preliminary anti-neoplastic activity of XMT-1536.

The secondary goals of DES and amplification studies are to evaluate the pharmacokinetics (PK) of XMT-1536, its release products and selected metabolites, to evaluate the development of anti-drug antibodies to XMT-1536, and to evaluate the effect of NaPi2b on XMT-1536. The relationship between tumor performance and objective tumor response.

The exploration goal of DES and amplification studies is to retrospectively assess the relationship between objective response and tumor manifestations of genes other than NaPi2b or other tumor molecular characteristics. Measuring effectiveness

In the two phases of the study, the evaluation based on computerized tomography (CT) scan and RECIST version 1.1 criteria was performed at the end of the second cycle and at the end of each even-numbered cycle thereafter. Patients are continuously evaluated for adverse events, use of concomitant medications, and infusion reactions in any cycle. Evaluate the pharmacokinetic profile of XMT-1536 and the development of selected release products/metabolites and anti-drug antibodies. RECIST version 1.1 was used to measure tumor response. Measure the objective response rate (ORR-complete response rate [CR] and partial response rate [PR]) and disease control rate (DCR-CR rate, PR rate and stable disease [SD]) of each team. The duration of response, progression-free survival (PFS) and overall survival (OS) were evaluated.

The main analysis of effectiveness is performed using data from both DES and EXP. The main indicator of ORR is the ORR obtained by the researcher’s radiological review, and is defined as the proportion of patients who have achieved PR or CR confirmed in accordance with RECIST 1.1. The secondary indicator is DCR, which is defined as the proportion of patients with complete response, partial response and/or stable disease in any duration according to RECIST 1.1. Summarize the number and percentage of patients who achieved response or clinical benefit, and provide an accurate 95% confidence interval (CI).

Whenever possible, the standard response criteria report includes analysis of duration of response (DOR), progression-free survival (PFS), and overall survival (OS). These statistical reports have a 95% CI median and quartile Kaplan-Meier (Kaplan-Meier) evaluation values.

The efficacy index is analyzed using both the efficacy analysis set and the efficacy evaluation analysis set of each cancer type team. Pharmacokinetic analysis

The PK profile of the active ingredients of XMT-1536, its release products and selected metabolites are determined for each patient using standard PK software (such as Phoenix WinNonlin) non-compartmental analysis. The PK parameters of each patient include the time of maximum observation concentration (t _max ), maximum concentration (C _max ), and the area under the concentration curve of the last measurable concentration (AUC _0-last ). When identifying the terminal elimination period, additional parameters such as elimination half-life, clearance rate, and volume of distribution are also determined.

PK analysis plan provides the missing concentration (missing concentration) and covariate data, outliers and values below the limit of quantification, and details of modeling the relationship between dose response and PK parameter response. Statistical Analysis

Prepare a statistical analysis plan (SAP), and the plan contains three parts: (1) from DES, (2) from EXP, and (3) from data analysis of the combined data of DES and EXP. This SAP system proposes the analysis of data recorded in clinical databases, laboratory data and other data. Each part is completed before the selection of the part is completed and the database of a specific research stage (ie, DES or EXP) is locked. The selected tables, lists, or graphs (collectively referred to as TLF) after the end of DES and EXP are generated for audit purposes and are indicated in SAP, but analysis reports are not prepared for these intermediate report values. Create a final analysis report and include it in the final clinical study. The final TLF and clinical research report is prepared after the data from the two phases have been locked. After the selection of this part has been completed, any changes made to the SAP of this part are shared with the researcher. Any deviations from the plan analysis are described in the final research report.

Transmit clinical data to PK supplier to support PK analysis. A separate analysis plan for the PK profile of XMT-1536, its released products and metabolites is prepared.

Before starting EXP administration, at a minimum, the data from DES should be appropriately summarized and reviewed with SRC. Based on the review of DES data, planning revisions can be created and submitted for approval before the start of the EXP phase. The statistical analysis is performed by Novella Clinical using SAS version 9.3 or higher.

Continuous variables including baseline characteristics are summarized by reporting the number of observations, average, standard deviation, median, minimum and maximum. The classification/discrete variables are summarized by the frequency table, which shows the number and percentage of patients in the classification. The time-to-event data is summarized using the Kaplan-Meyer method.

Unless otherwise instructed, summarized statistics are reported only with observed data. Does not estimate missing data. If the baseline value is missing, the change from the baseline is not calculated. Baseline is defined as the last available observation value before the first administration of study drug on the first cycle. The disposal of lost data is specified in SAP and the method used to report the endpoint. result

As of February 2019, 20 patients have crossed 6 dose levels (3 mg/m ² to 40 mg/m ² ) in a 21-day cycle and 16 patients have crossed 2 dose levels (20 mg/m ² ) in a 28-day cycle. /m ² to 30 mg/m ² ) administration. Thirteen of the 20 patients completed the 21-day DLT evaluation period. Sixteen of the 16 patients completed the 28-day evaluation period. One DLT attributable to study drug occurred in patients who were dosed at 40 mg/m ² in a 21-day cycle and one DLT attributable to study drug occurred in patients who were dosed at 30 mg/m ² in a 28-day cycle DLT. Two occurs and Study drug related SAE: the DL 4,2 mg / m ^2, congestive heart failure, and a 21 day cycle in heat DL 5,30 mg / m ^2, 21-day cycle. Treatment-related adverse events (TRAE) are grade 1 or 2; the most common TRAEs so far are anemia, joint pain, fatigue, vomiting, anorexia, increased AST, diarrhea, dyspnea (irrelevant or unlikely to be related), low whiteness Proteinemia (irrelevant or unlikely to be related), hypoxia (irrelevant), headache, nausea, proteinuria, and vomiting. 10 patients had been re-staged after treatment, and 9 patients had stable disease (SD) as the best response.

As of August 2019, 51 patients have crossed 9 dose levels (3 mg/m ² to 40 mg/m ² (n=20) for the 21-day regimen and 20 mg/m ² to 36 mg/m ^{2 for the} 28-day regimen. m ² (n=31) administration). One DLT occurred in one patient who was dosed at 30 mg/m ² in a 28-day cycle and one DLT occurred in one patient who was dosed at 36 mg/m ² in a 28-day cycle. Both are believed to be related to XMT-1536.

Table 7 presents the characteristics of patients enrolled with a 21-day cycle across 6 dose levels (2 to 40 mg/m ² ) and a 28-day cycle across 2 dose levels (20 to 30 mg/m ² ). Table 7 Dose level Dose (mg/m ² ) Cycle duration (days) Tumor type N Age range Y/o Shortest/longest study duration 1 3 twenty one N=1 ovary 55 4 cycles 2 6 twenty one N=1 ovary 54 2 cycles 3 12 twenty one N=7 Ovary-1 NSCLC-2 Endometrium-3 Papillary kidney-1 39 to 67 average=54 median=70 1 to 4 cycles 4 20 twenty one N=6 Ovary-2 NSCLC-1 Endometrium-1 Fallopian tube-1 Salivary gland duct-1 58 to 94 average=72 median=70 1 to 10 cycles 5 30 twenty one N=4 Ovary-3 NSCLC-1 63 to 68 average=66 median=66 1 to 4 cycles 6 40 twenty one N=1 ovaries are selected 54 1 cycle 4A 20 28 N=8 ovary N=1 papillary kidney 47-74 Average=62 Median=63 1 to 6 cycles 5A 30 28 N=7 51-76 average=68 median=71 2 to 3 cycles

The plan was revised in August 2018. The 21-day cycle was extended to 28 days and the observation period of the DLT assessment was extended to include two 28-day cycles. This is the period after the safety review committee assesses the patient's response to XMT-1536 and decides whether a dose escalation should occur. Table 8 is a summary of the medications administered to patients as of February 2019. Table 8 Dose level (DL) Dosing regimen Number of patients DL 1, 3 mg/m ² 21-day cycle, cycle 1 of the DLT observation period 1 DL 2, 6 mg/m ² 21-day cycle, cycle 1 of the DLT observation period 1 DL 3, 12 mg/m ² 21-day cycle, cycle 1 of the DLT observation period 7 DL 4, 20 mg/m ² 21-day cycle, cycle 1 of the DLT observation period 6 DL 5, 30 mg/m ² 21-day cycle, cycle 1 of the DLT observation period 4 DL 6, 40 mg/m ² 21-day cycle, cycle 1 of the DLT observation period 1 DL 4A, 20 mg/m ² 28-day cycle, the second cycle of the DLT observation period 9 DL 5A, 30 mg/m ² 28-day cycle, the second cycle of the DLT observation period 7

Figure 1 is a "swim lane chart" that summarizes the details of the study time of 20 patients (n=19) in a 21-day cycle and 4 patients (n=4) administered in a 28-day cycle through DL6. The graph is plotted on the y-axis by the patient's dose level and tumor type relative to the study drug time on the x-axis. The x-axis arranges weekly intervals. At lower doses, most patients withdrew from the study because of progressive disease, but most patients had stable disease starting at DL4.

Figure 2 is a "waterfall chart", which shows the best response to patients who have been administered at least one scan so far based on the RECIST criteria. The y-axis shows the percentage change in tumor size. The x-axis arranges the dose level response. Although the number of patients at each dose is small, there seems to be a dose response. Two ongoing patients almost achieved partial remission starting at DL 5 (defined as a reduction in tumor size of more than or equal to 30% since baseline).

Figure 3 shows the overall best response of 11 patients, as well as the H score and CA-125 levels of ovarian cancer patients administered in a 21-day cycle.

Figure 4 is a "waterfall chart", which shows the best response to patients who have been administered at least one scan in a 21-day cycle and a 28-day cycle according to the RECIST criteria. The y-axis shows the percentage change in tumor size. The x-axis arranges the dose level response. Although the number of patients at each dose is small, there seems to be a dose response. Two patients almost achieved partial remission starting at DL 5 (defined as a reduction of more than or equal to 30% in tumor size from baseline). One patient achieved partial remission at DL 4A.

Figure 5 is a "lane chart" that summarizes the details of the study time of patients who passed DL6, 21-day cycle (n=20) and DL 4A and DL 5A patients (n=16) administered in 28-day cycle. The graph is plotted on the y-axis by the patient's dose level and tumor type relative to the study drug time on the x-axis. The x-axis arranges weekly intervals. At lower doses, most patients withdrew from the study because of progressive disease, but most patients had stable disease starting at DL4.

As of May 10, 2019, 37 patients have crossed 6 dose levels (3 mg/m ² to 40 mg/m ² ) in a 21-day cycle and 17 patients have crossed 2 dose levels in a 28-day cycle ( 20 mg/m ² to 30 mg/m ² ) administration. Table 9 summarizes the patient characteristics. The treatment was well tolerated, and most adverse events were reported as grade 1 or 2 and no grade 4 or 5 TRAEs. The low rates of toxicity often associated with microtubule targeting agents or ADCs, such as neutropenia, ocular toxicity, or peripheral neuropathy, are observed.

Table 10 summarizes TRAE in ≥10% of patients. Report 13 patients (33%) serious adverse events (SAE). Two SAEs that may be related to treatment are grade 2 fever at DL5 (30 mg/m ² ) and congestive heart failure at grade 3 DL4 (20 mg/m ² ). Seventeen SAEs that were not or unlikely to be related to treatment occurred in 11 patients: bowel/small bowel obstruction (3), disease progression (2), hypoxia (2), pleural effusion (2), abdominal pain, acute Hemorrhagic anemia, cellulitis Staphylococcus, cerebrovascular accident, hemorrhagic anemia, malignant ascites, pericardial effusion, subdural hematoma and vaginal bleeding. Report two dose-limiting toxicities in DL5A (30 mg/m ² ) and DL6 (40 mg/m ² ). Under DL5A, on the 8th day of the first cycle, the level 3 AST is raised, and the level 2 is returned within 7 days and the level 1 is returned within 13 days. An increase in AST was accompanied by an increase in grade 1 alkaline phosphatase, but there were no reports of an increase in ALT or bilirubin. Under DL6, on the 8th day of the 1st cycle, the level 3 AST is raised, and the level 1 is returned within 21 days. The increase in AST was accompanied by an increase in ALT of grade 2 (which subsided to grade 1 within 8 days) and an increase in alkaline phosphatase of grade 1, but there were no reports of increased bilirubin. Table 9 Patient characteristics (N=37) age) Median (range) 64(39-93) Gender-N (%) Female male 32(86) 5(14) ECOG performance status-N (%) 0 1 11(30) 26(70) Tumor type-N (%) Ovary, fallopian tube or primary peritoneum NSCLC Endometrial papillary renal salivary gland duct 22(59) 4(11) 8(22) 2(5) 1(3) Previous lines of treatment for metastatic disease Median (range) 4(1-13) Previous treatment line, only ovarian cancer (N=22) Median (range) 5(1-11) Table 10 Better project Grade 1 Level 2 Level 3 total nausea 12(32) 2(5) 0 14(38) fatigue 4(11) 7(19) 0 11(30) headache 5(14) 5(14) 0 10(27) Increased aspartate transaminase (AST) 3(8) 2(5) 4(11) 9(24) Reduce appetite 1(3) 6(16) 0 7(19) Increased blood alkaline phosphatase 6(16) 0 0 6(16) Vomiting 4(11) 1(3) 0 5(14) Increased γ-glutamine converting enzyme (GGT) 3(8) 1(3) 0 4(11) Myalgia 3(8) 1(3) 0 4(11) fever 3(8) 1(3) 0 4(10)

The plasma PK profile of the analytes (total antibody, conjugated drug (AF-HPA), free drug (AF-HPA) and drug metabolite auristatin F (AF)) was determined in blood samples from 30 patients ( 3 mg/m ² , n=1; 6 mg/m ² , n=1; 12 mg/m ² , n=7; 20 mg/m ² , n=15; 30 mg/m ² , n=5; 40 mg/m ² , n=1). The results normalized by the dose showed that the exposure increased with the increase of the dose and was almost proportional to the dose. The PK characteristics are consistent with those reported by other approved antibody-drug conjugates or those currently in clinical development. Compared with conjugated drugs, free drugs (AF-HPA) or its metabolites (AF) have lower systemic exposure. In addition, after multiple doses, no free drug or its metabolites accumulated.

If feasible, submit slices or whole tissue blocks from the patient's tumor for IHC performance analysis. The Department of Immunohistochemistry is established using automated TechMate 500 or TechMate 1000 (BioTek Solutions/Ventana medical Systems) platforms, where various antigen repair conditions and primary antibody titers are tested to develop higher throughput plans. The plan is selected based on the staining of control materials and the staining of preclinical materials that have a known response to ADC treatment. In short, the 4 µ slices used to create the plan are cut, thawed and rehydrated with xylene and a series of alcohols. Put the slices in a standard steamer and use BioGenex AR-10 repair solution for antigen repair. Perform further repairs on the TechMate platform with proteinase K (DAKO). After serum blocking: administer the primary antibody (anti-NaPi2b/MERS67) at room temperature for 30 minutes, then use endogenous peroxidase blocking solution and non-biotin polymer detection (rabbit Polink-2 Plus detection System, GBI), and finally a brief counterstaining with hematoxylin.

Tumor membrane staining was evaluated by manual reading using the "H-score" method on a light microscope. The sections stained with hematoxylin and eosin and the sections stained with the rabbit IgG isotype control are evaluated as part of the scoring process. The same scoring method was used for all tumor types evaluated. The H-score incorporates the staining intensity (intensity measurement increased from 0 to 3+) and the percentage of positive cells detected on the tumor cell membrane. H-score=((% of <1+)X 0)+((% of 1+)X 1)+((% of 2+)X 2)+((% of 3+)X 3). Samples that do not show too few cells or those that do not show primary tumors in the submitted samples.

Figure 6 shows NaPi2b protein expression in 34 patient tumor types (eg, ovary, NSCLC, endometrium, papillary kidney, and salivary gland ducts), where fallopian tube and primary peritoneal tumor types are combined with ovarian tumor types. The y-axis is the number of positive tumor cells in the NaPi2b IHC assay and the H score of the staining intensity, ranging from 0 to 300. Detectable NaPi2b protein expression was observed in all samples of ovarian cancer and lung adenocarcinoma.

Figure 7 is a "lane chart" which summarizes the details of the study time of patients who passed DL6, 21-day cycle (n=20) and DL4A and DL5A patients (n=17) administered in 28-day cycle. The graph is plotted on the y-axis by the patient's dose level and tumor type relative to the study drug time on the x-axis. The x-axis arranges weekly intervals. At lower doses, most patients withdrew from the study because of progressive disease, but most patients had stable disease starting at DL4.

Table 11 summarizes the reasons for discontinuing participation in studies that cross dose levels. Table 12 shows the result response of the evaluable group. Based on the objective response and duration of treatment, clinical activity was observed at a dose of 20 mg/m ² or higher. Table 11 DL1 3 mg/m ² n=1 DL2 6 mg/m ² n=1 DL3 12 mg/m ² n=7 DL4 & 4A 20 mg/m ² n=13 DL5 & 5A 30 mg/m ² n=11 DL6 40 mg/m ² n=1 Total N=34 ^b Progressive diseases according to RECIST 1 1 3 7 6 18 (53%) ^A clinical progression 4 2 3 9 (26%) Patient selection 2 1 1 4 (12%) Physician choice 2 1 3 (9%) ^a 3 out of 10 patients died, not related to XMT-1536 ^b 3 patients in progress Table 12 result All ovarian cancer (OC) All NSCLC OC ≥20 mg/m ² NSCLC ≥20 mg/m ² OC + NSCLC≥20 mg/m ² OC ≥30 mg/m ² N 19 3 16 2 18 7 PR ^* 3 (16%) 0(0%) 3 (19%) 0(0%) 3 (17%) 2 (28%) SD ^* 8 (42%) 2 (67%) 6 (38%) 2(100%) 8 (44%) 3 (43%) DCR (PR + SD) 11 (58%) 2 (67%) 9 (57%) 2(100%) 11 (61%) 5 (71%) Duration of treatment> 16 weeks 8 (42%) 1 (33%) 9 (57%) 1 (50%) 10 (56%) 3 (43%) PD ^* 8 (42%) 1 (33%) 7 (43%) 0(0%) 7 (39%) 2 (28%) *Measured with RECIST version 1.1

A 70-year-old woman with platinum drug-resistant high-grade serous ovarian cancer and 11 lines of previous treatment was treated with DL4A (20 mg/m ² ). The target lesions of 52 and 42 mm around the liver and mid-abdominal metastases were reduced by 40% at the end of the second cycle (four-week cycle) and 75% at the end of the third cycle. Objective responses were observed at doses of 20 mg/m ² and higher. Among patients with ovarian cancer and lung adenocarcinoma treated with ≥20 mg/m ² (N=18), 3 (17%) PR and 8 (44%) SD (ORR 17%, SD 44%) have been observed , DCR 61%), in 9 patients (50%), the duration of treatment at doses of 20 mg/m ² and higher was> 16 weeks (n=18).

Table 13 shows the characteristics of selected patients who crossed 7 dose levels (2 to 40 mg/m ² ), 21-day cycle and 3 dose levels (20 to 36 mg/m ² ), 28-day cycle as of August 2019 . Table 13 Dose level Dose (mg/m ² ) Cycle duration (days) Tumor type N Age range Y/o Shortest/longest study duration 1 3 twenty one N=1 ovary 55 4 cycles 2 6 twenty one N=1 ovary 54 2 cycles 3 12 twenty one N=7 Ovary-1 NSCLC-2 Endometrium-3 Papillary kidney-1 39 to 67 average=54 median=70 1 to 4 cycles 4 20 twenty one N=6 Ovary-2 NSCLC-1 Endometrium-1 Fallopian tube-1 Salivary gland duct-1 58 to 94 average=72 median=70 1 to 10 cycles 5 30 twenty one N=4 Ovary-3 NSCLC-1 63 to 68 average=66 median=66 1 to 4 cycles 6 40 twenty one N=1 ovaries are selected 54 1 cycle 4A 20 28 N=8 ovary N=1 papillary kidney 47 to 74 average=62 median=63 1 to 12 cycles 5A 30 28 N=8 Ovary-4 Endometrium-4 51 to 76 Average=68 Median=71 2 to 6 cycles Additional 5A 30 28 N=7 Ovary-6 NSCLC-1 51 to 85 average=68 median=71 2 cycles 6A 28 N=7 Ovary-5 NSCLC-2 47 to 63 average=54 median=56 2 cycles

The plan was revised in July 2019. Join the eligibility criteria for ovarian cancer and non-small cell lung cancer patients. Table 14 is a summary of the patients who were administered as of August 2019. Table 14 Dose level (DL) Dosing regimen Number of patients DL 1, 3 mg/m ² 21-day cycle, cycle 1 of the DLT observation period 1 DL 2, 6 mg/m ² 21-day cycle, cycle 1 of the DLT observation period 1 DL 3, 12 mg/m ² 21-day cycle, cycle 1 of the DLT observation period 7 DL 4, 20 mg/m ² 21-day cycle, cycle 1 of the DLT observation period 6 DL 5, 30 mg/m ² 21-day cycle, cycle 1 of the DLT observation period 4 DL 6, 40 mg/m ² 21-day cycle, cycle 1 of the DLT observation period 1 DL 4A, 20 mg/m ² 28-day cycle, the second cycle of the DLT observation period 9 DL 5A, 30 mg/m ² 28-day cycle, the second cycle of the DLT observation period 8 DL 5A, 30 mg/m ² , additional 28-day cycle, the second cycle of the DLT observation period 7 DL 6A, 36 mg/m ² 28-day cycle, the second cycle of the DLT observation period 7 Figure 8 is a "swimming lane diagram" that summarizes the patients across 7 dose levels (2 to 40 mg/m ² ), a 21-day cycle, and 3 dose levels (20 to 36 mg/m ² ), a 28-day cycle Details of the study time. The graph is plotted on the y-axis by the patient's dose level and tumor type relative to the study drug time on the x-axis. The x-axis arranges weekly intervals. At lower doses, most patients withdrew from the study because of progressive disease, but most patients had stable disease starting at DL4. The longest study duration for patients in the ongoing trial is 50 weeks. in conclusion

XMT-1536 has well tolerated dose levels up to 30 mg/m ² or 36 mg/m ² or 43 mg/m ² and has early signs of anti-tumor activity. MTD or RP2D has not been identified. Dose escalation continues in patients with NaPi2b-expressing ovarian cancer, NSCLC, papillary thyroid cancer, endometrial cancer, papillary renal cell carcinoma, and salivary duct cancer. other instructions

Although the present invention has been described together with its detailed description, the foregoing description is intended to exemplify rather than limit the scope of the invention defined by the appended claims. Other aspects, advantages and modifications are within the scope of the following patent applications.

[Figure 1] is a swimmers plot, which summarizes the details of the study time of all patients (n=19) through DL6, 21-day cycle, and four patients (n=4) who were administered the 28-day cycle .

[Figure 2] is a "waterfall plot", which shows the best response to patients from DL1 to DL5 in the 21-day treatment cycle and who have performed at least one scan so far according to the RECIST criteria.

[Figure 3] shows the best overall response of all patients (n=11) in the 21-day treatment cycle.

[Figure 3] shows the best overall response of all patients (n=11) in the 21-day treatment cycle.

[Figure 4] is a "waterfall chart", which shows the best response to patients who have been administered at least one scan in a 21-day cycle and a 28-day cycle according to the RECIST guidelines.

[Figure 5] is a "lane chart" that summarizes the details of the study time of all patients who passed DL6, 21-day cycle (n=20) and DL 4A and DL 5A patients (n=16) administered in 28-day cycle .

[Figure 6] shows the H-scores obtained by IHC NaPi2b protein expression in tumor samples from 34 patients. The y-axis shows the H-score of NaPi2b and the x-axis shows the tumor type.

[Figure 7] is a "lane chart", which summarizes the details of the study time of all patients who passed DL6, 21-day cycle (n=20) and DL 4A and DL 5A patients (n=17) administered in 28-day cycle .

[Figure 8] is a "lane diagram" that summarizes the 7 dose levels (2 to 40 mg/m ² ), a 21-day cycle and 3 dose levels (20 to 36 mg/m ² ), a 28-day cycle Details of the patient's study time.

Claims (40)

A method for treating NaPi2b-expressing tumors in individuals in need thereof, which method comprises infusion of approximately 10 mg/m ² to 43 mg/m ² in a dose of approximately between 10 mg/m ² and 43 mg/m ² on the first day of treatment and every four weeks thereafter The individual administers an antibody polymer-drug conjugate targeting NaPi2b, wherein the conjugate comprises a NaPi2b antibody, and the NaPi2b antibody comprises: CDRH1 comprising the amino acid sequence GYTFTGYNIH (SEQ ID NO: 5); comprising the amino acid CDRH2 of the sequence AIYPGNGDTSYKQKFRG (SEQ ID NO: 6); CDRH3 including the amino acid sequence GETARATFAY (SEQ ID NO: 7); CDRL1 including the amino acid sequence SASQDIGNFLN (SEQ ID NO: 8); including the amino acid sequence YTSSLYS (SEQ ID NO: 9) CDRL2; and CDRL3 comprising the amino acid sequence QQYSKLPLT (SEQ ID NO: 10); and the polymer-drug conjugate of formula A:

Wherein: The polymer comprises poly(1-hydroxymethylethylene hydroxymethyl-formal) (PHF) with a molecular weight ranging from about 5 kDa to about 10 kDa; m is an integer from 20 to 75, and m ₁ is An integer from about 5 to about 35, m ₂ is an integer from about 3 to about 10, m _3a is an integer from 0 to about 4, m _3b is an integer from 1 to about 5, m, m ₁ , m ₂ , m _3a and The sum of m _3b is in the range of about 40 to about 75, and m ₅ is an integer of about 2 to about 5. According to the method of claim 1, wherein the NaPi2b antibody comprises a variable heavy chain and a variable light chain, the variable heavy chain comprises the amino acid sequence of SEQ ID NO: 3, and the variable light chain comprises SEQ ID NO: 4 amino acid sequence. According to the method of claim 1, wherein the NaPi2b antibody comprises a heavy chain and a light chain, the heavy chain comprises the amino acid sequence of SEQ ID NO: 1, and the light chain comprises the amino acid of SEQ ID NO: 2 sequence. According to the method of items 1 to 3 in the scope of patent application, the dosage is about 11.5 mg/m ² to about 12.5 mg/m ² . The method according to any one of items 1 to 3 in the scope of patent application, wherein the dosage is about 12 mg/m ² . The method according to any one of items 1 to 3 in the scope of patent application, wherein the dosage is about 19.5 mg/m ² to about 20.5 mg/m ² . According to the method of items 1 to 3 in the scope of patent application, the dosage is about 20 mg/m ² . The method according to any one of items 1 to 3 in the scope of patent application, wherein the dosage is about 24.5 mg/m ² to about 26.5 mg/m ² . According to the method of items 1 to 3 in the scope of patent application, the dosage is about 25 mg/m ² . The method according to any one of items 1 to 3 in the scope of the patent application, wherein the dosage is about 29.5 mg/m ² to about 31.5 mg/m ² . According to the method of items 1 to 3 in the scope of patent application, the dosage is about 30 mg/m ² . The method according to any one of items 1 to 3 in the scope of patent application, wherein the dosage is about 32.5 mg/m ² to about 33.5 mg/m ² . According to the method of items 1 to 3 in the scope of patent application, the dosage is about 33 mg/m ² . The method according to any one of items 1 to 3 in the scope of the patent application, wherein the dosage is about 35.5 mg/m ² to about 36.5 mg/m ² . According to the method of items 1 to 3 in the scope of patent application, the dosage is about 36 mg/m ² . The method according to any one of items 1 to 3 in the scope of patent application, wherein the dose is about 42.5 mg/m ² to about 43.5 mg/m ² . According to the method of items 1 to 3 in the scope of patent application, the dosage is about 43 mg/m ² . The method according to any one of items 1 to 3 in the scope of patent application, wherein the dosage is about 10 mg/m ² , 15 mg/m ² , 20 mg/m ² , 25 mg/m ² , 30 mg/m ² , 33 mg/m ² , 36 mg/m ² , 43 mg/m ² or 45 mg/m ² . The method according to any one of the aforementioned patent applications, wherein the individual is a human. The method according to any one of the aforementioned patent applications, wherein the tumor is selected from the group consisting of: ovarian cancer, non-small cell lung cancer (NSCLC), papillary thyroid cancer, endometrial cancer, mastoid Renal cell carcinoma, cholangiocarcinoma, salivary duct cancer, clear cell renal cancer, breast cancer, kidney cancer, or cervical cancer. According to the method according to item 20 of the scope of patent application, the ovarian cancer is epithelial ovarian cancer, fallopian tube cancer or primary peritoneal cancer. According to the method of item 21 in the scope of patent application, the epithelial ovarian cancer is classified into high-grade serous ovarian cancer, low-grade ovarian cancer or clear cell ovarian cancer. According to the method according to items 16 to 22 of the scope of patent application, the ovarian cancer is resistant to platinum, and the individual has received no more than 3 lines of previous treatment. According to the method of items 16 to 23 in the scope of patent application, the individual has ovarian cancer and has received no more than 3 lines of previous treatment. According to the method of item 20 of the scope of patent application, the NSCLC cancer is non-squamous. According to the method of item 20 in the scope of patent application, the NSCLC cancer is subclassified as adenocarcinoma. According to the method according to item 25 or 26 of the scope of patent application, the individual has NSCLC and has received previous treatment with a platinum-based regimen and PD-1 or PD-L1 monoclonal antibody. According to the method of item 27 in the scope of patent application, the platinum-based regimen and PD-1 or PD-L1 monoclonal antibody system are administered in combination. According to the method of item 27 of the scope of patent application, the platinum-based solution and the PD-1 or PD-L1 monoclonal antibody system are administered sequentially. According to the method of items 27 to 29 in the scope of patent application, the platinum-based solution is cisplatin or carboplatin. According to the method of items 27 to 30 in the scope of patent application, the individual has not received cytotoxic agent treatment or immunotherapy treatment. According to the method of items 27 to 31 in the scope of the patent application, the system uses known carcinogenic mutations to prove intolerance to approved treatments or disease progression. According to the method according to item 20 of the scope of patent application, the papillary thyroid cancer is a progressive, radioiodine refractory, locally recurring or metastatic disease. According to the method of item 33 in the scope of patent application, the individual has papillary thyroid cancer and is resistant or intolerant to kinase inhibitor therapy. According to the method of item 20 of the scope of patent application, the endometrial cancer is epithelial endometrial cancer. According to the method of item 20 or 35 of the scope of patent application, the endometrial cancer is not a stromal tumor or carcinosarcoma. The method according to item 20, 35 or 36 of the scope of patent application, wherein the individual has endometrial cancer and has received previous treatment with carboplatin/paclitaxel or a similar regimen. The method according to item 20, 35 or 36 of the scope of patent application, wherein the individual has endometrial cancer and has received previous treatment for low-grade hormone receptor positive endometrial adenocarcinoma. According to the method of item 20 in the scope of patent application, the papillary renal cell carcinoma has a significant papillary growth pattern. According to the method of item 20 of the scope of patent application, the cancer has a histological diagnosis of salivary duct carcinoma that has progressed after standard systemic treatment.

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